Although lipid analyst tend to have a firm understanding of what is meant by the term "lipid", there is no widely-accepted definition.

General text books usually describe lipids in woolly terms as a group of naturally occurring compounds, which have in common a ready solubility in such organic solvents as hydrocarbons, chloroform, benzene, ethers and alcohols. They include a diverse range of compounds, like fatty acids and their derivatives, carotenoids, terpenes, steroids and bile acids. It should be apparent that many of these compounds have little by way of structure or function to relate them. In fact, a definition of this kind is positively misleading, since many of the substances that are now widely regarded as lipids may be almost as soluble in water as in organic solvents.

While the international bodies that usually decide such matters have shirked the task, a more specific definition of lipids than one based simply on solubility is necessary, and most scientist active in this field would happily restrict the use of "lipid" to fatty acids and their naturally-occurring derivatives (esters or amides). The definition could be stretched to include compounds related closely to fatty acid derivatives through biosynthetic pathways (e.g. aliphatic ethers or alcohols) or by their biochemical or functional properties (e.g. cholesterol) . My definition is Lipids are fatty acids and their derivatives, and substances related biosynthetically or functionally to these compounds. This treats cholesterol (and plant sterols) as a lipid, and could be interpreted to include bile acids, tocopherols and certain other compounds. It also enables classification of such compounds as gangliosides as lipids, although they are more soluble in water than in organic solvents. However, it does not include such natural substances as steroidal hormones, petroleum products, some fat-soluble vitamins, most polyketides, carotenoids or simple terpenes, except in rare circumstances. If "lipids" are defined in this way, fatty acids must be defined also. They are compounds synthesised in nature via condensation of malonyl coenzyme A units by a fatty acid synthase complex. They usually contain even numbers of carbon atoms in straight chains (commonly C14 to C24), and may be saturated or unsaturated; they can also contain other substituent groups. Fahy et al. (J. Lipid Res., 46, 839-862 (2005)) have developed a classification system for lipids that holds promise (see our page on Nomenclature). While their definition of a lipid is too broad for my taste, it is based on sound scientific principles (although these may not mean much to non-biochemists), i.e.

Lipids are hydrophobic or amphipathic small molecules that may originate entirely or in part by carbanion-based condensations of thioesters (fatty acids, polyketides, etc.) and/or by carbocation-based condensations of isoprene units (prenols, sterols, etc.). The most common lipid classes in nature consist of fatty acids linked by an ester bond to the trihydric alcohol - glycerol, or to other alcohols such as cholesterol, or by amide bonds to sphingoid bases, or on occasion to other amines. In addition, they may contain alkyl moieties other than fatty acids, phosphoric acid, organic bases, carbohydrates and many more components, which can be released by various hydrolytic procedures. A further subdivision into two broad classes is convenient for chromatography purposes especially. Simple lipids are defined as those that on hydrolysis yield at most two types of primary product per mole; complex lipids yield three or more primary hydrolysis products per mole. Alternatively, the terms "neutral" and "polar" lipids respectively are used to define these groups, but are less exact. The complex lipids for many purposes are best considered in terms of either the glycerophospholipids (or simply if less accurately as phospholipids), which contain a polar phosphorus moiety and a glycerol backbone, or the glycolipids (both glycoglycerolipids and glycosphingolipids), which contain a polar carbohydrate moiety, as these are more easily analysed separately. The picture is further complicated by the existence of phosphoglycolipids and sphingophospholipids (e.g. sphingomyelin). See also our page on Lipid Nomenclature.

Simple Lipids
Triacylglycerols: Nearly all the commercially important fats and oils of animal and plant origin consist almost exclusively of the simple lipid class triacylglycerols (termed "triglycerides" in the older literature). They consist of a glycerol moiety with each hydroxyl group esterified to a fatty acid. In nature, they are synthesised by enzyme systems, which determine that a centre of asymmetry is created about carbon-2 of the glycerol backbone, so they exist in enantiomeric forms, i.e. with different fatty acids in each position.

A stereospecific numbering system has been recommended to describe these forms. In a Fischer projection of a natural L-glycerol derivative, the secondary hydroxyl group is shown to the left of C-2; the carbon atom above this then becomes C-1 and that below is C-3. The prefix "sn" is placed before the stem name of the compound, when the stereochemistry is defined. Their primary biological function is to serve as a store of energy. As an example, the single molecular species 1,2-dihexadecanoyl-3-(9Zoctadecenoyl)-sn-glycerol is illustrated. (More...).

Diacylglycerols (less accurately termed "diglycerides") and monoacylglycerols (monoglycerides) contain two moles and one mole of fatty acids per mole of glycerol, respectively, and exist in various isomeric forms. They are sometimes termed collectively "partial glycerides". Although they are rarely present at greater than trace levels in fresh animal and plant tissues, 1,2-diacyl-sn-glycerols are key intermediates in the biosynthesis of triacylglycerols and other lipids, and they are vital cellular messengers, generated on hydrolysis of phosphatidylinositol and related lipids by a specific phospholipase C. Synthetic materials have importance in commerce. (More..).

2-Monoacyl-sn-glycerols are formed as intermediates or end-products of the enzymatic hydrolysis of triacylglycerols; these and other positional isomers are powerful surfactants. 2-Arachidonoylglycerol has important biological properties (as an endocannabinoid). Acyl migration occurs rapidly in partial glycerides at room temperature, but especially on heating, in alcoholic solvents or in the presence of acid or base, so special procedures are required for their isolation or analysis if the stereochemistry is to be retained. Synthetic 1/3-monoacylglycerols are important in commerce as surfactants. (More..).

Sterols and sterol esters: Cholesterol is by far the most common member of a group of steroids in animal tissues; it has a tetracyclic ring system with a double bond in one of the rings and one free hydroxyl group. It is found both in the free state, where it has an essential role in maintaining membrane fluidity, and in esterified form, i.e. as cholesterol esters. Other sterols are present in free and esterified

form in animal tissues, but at trace levels only. Cholesterol is the precursor of the bile acids and steroidal hormones. In plants, cholesterol is rarely present in other than small amounts, but such phytosterols as sitosterol, stigmasterol, avenasterol, campesterol and brassicasterol, and their fatty acid esters are usually found, and they perform a similar function. (More..). Hopanoids are related lipids produced by some bacterial species. Waxes: In their most common form, wax esters consist of fatty acids esterified to longchain alcohols with similar chain-lengths. The latter tend to be saturated or have one double bond only. Such compounds are found in animal, plant and microbial tissues and they have a variety of functions, such as acting as energy stores, waterproofing and lubrication.

In some tissues, such as skin, avian preen glands or plant leaf surfaces, the wax components can be much more complicated in their structures and compositions. They can contain aliphatic diols, free alcohols, hydrocarbons (e.g. squalene), aldehydes and ketones. (More..).

Tocopherols (collectively termed vitamin E) are substituted benzopyranols (methyl tocols) that occur in vegetable oils. Different forms (-, -, - and -) are recognized according to the number or position of methyl groups on the aromatic ring. -Tocopherol (with the greatest Vitamin E activity) illustrated is an important natural antioxidant. Tocotrienols have similar ring structures but with three double bonds in the aliphatic chain. (More...). Free (unesterified) fatty acids are minor constituents of living tissues but are of biological importance as precursors of lipids as an energy source and as cellular messengers. (More...).

Glycerophospholipids
Phosphatidic acid or 1,2-diacyl-sn-glycerol-3-phosphate is found in trace amounts only in tissues under normal circumstances, but it has great metabolic importance as a

biosynthetic precursor of all other glycerolipids. It is strongly acidic and is usually isolated as a mixed salt. One specific isomer is illustrated as an example.

Lysophosphatidic acid with one mole of fatty acid per mole of lipid (in position sn-1) is a marker for ovarian cancer, and is a key cellular messenger. (More...).

Phosphatidylglycerol or 1,2-diacyl-sn-glycerol-3phosphoryl-1'-sn-glycerol tends to be a trace constituent of most tissues, but it is often the main component of some bacterial membranes. It has important functions in lung surfactant, where its physical properties are significant, and in plant chloroplasts, where it appears to have an essential role in photosynthesis. Also, it is the biosynthetic precursor of cardiolipin. In some bacterial species, the 3'-hydroxyl of the phosphatidylglycerol moiety is linked to an amino acid (lysine, ornithine or alanine) to form an Oaminoacylphosphatidylglycerol or complex 'lipoamino acid'. (More..). Cardiolipin (diphosphatidylglycerol or more precisely 1,3-bis(sn-3'-phosphatidyl)-snglycerol) is a unique phospholipid with in essence a dimeric structure, having four acyl groups and potentially carrying two negative charges (and is thus an acidic lipid). It is an important constituent of mitochondrial lipids especially, so heart muscle is a rich source. Amongst other functions, it plays a key role in modifying the activities of the enzymes concerned with oxidative phosphorylation. (More...).

Lysobisphosphatidic acid or bis(monoacylglycerol)phosphate is an interesting lipid as its stereochemical configuration differs from that of all other animal glycero-phospholipids in that the phosphodiester moiety is linked to positions sn-1 and sn-1' of glycerol, rather than to position sn-3 to which the fatty acids are esterified (some experts think that position sn-2 is more likely for the latter). It is usually a rather minor component of animal tissues, but is enriched in the lysosomal membranes of liver and appears to be a marker for this organelle. The glycerophosphate backbone is particularly stable, presumably because of the unusual stereochemistry. (More..).

Phosphatidylcholine or 1,2-diacyl-sn-glycerol3-phosphorylcholine (or "lecithin", although the term is now used more often for the mixed phospholipid by-products of seed oil refining) is usually the most abundant lipid in the membranes of animal tissues, and it is often a major lipid component of plant membranes, but only rarely of bacteria. With the other choline-containing phospholipid, sphingomyelin, it is a key structural component and constitutes much of the lipid in the external monolayer of the plasma membrane of animal cells especially. (More...). Lysophosphatidylcholine, which contains only one fatty acid moiety in each molecule, generally in position sn-1, is sometimes present as a minor component of tissues. It is a powerful surfactant and is more soluble in water than most other lipids.

Phosphatidylethanolamine (once given the trivial name "cephalin") is usually the second most abundant phospholipid class in animal and plant tissues, and can be the major lipid class in microorganisms. As part of an important cellular process, the amine group can be methylated enzymically to yield first phosphatidyl-N-monomethylethanolamine and then phosphatidyl-N,Ndimethylethanolamine, but these never accumulate in significant amounts; the eventual product is phosphatidylcholine. N-Acylphosphatidylethanolamine is a minor component of some plant tissues, especially cereals, and it is occasionally found in animal tissues, where it is the precursor of some biologically active amides. Lysophosphatidylethanolamine contains only one mole of fatty acid per mole of lipid. (More...).

Phosphatidylserine is a weakly acidic lipid that is present in most tissues of animals and plants and is also found in microorganisms. It is located entirely on the inner monolayer surface of the plasma membrane and other cellular membranes. Phosphatidylserine is an essential cofactor for the activation of protein kinase C, and it is involved in many other biological processes, including blood coagulation and apoptosis (programmed cell death). (More...). N-Acylphosphatidylserine has been detected in some animal tissues.

Phosphatidylinositol, containing the optically inactive form of inositol, myo-inositol, is a common constituent of animal, plant and microbial lipids. In animal tissues especially, it may be accompanied by small amounts of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate (and other 'poly-phosphoinositides'). These compounds have a rapid rate of metabolism in animal cells, and are converted to metabolites such as diacylglycerols and inositol phosphates, which are important in regulating vital processes. For example, diacylglycerols regulate the activity of a group of enzymes known as protein kinase C, which in turn control many

key cellular functions, including differentiation, proliferation, metabolism and apoptosis (see our web pages on phosphatidylinositol). In addition, phosphatidylinositol is the primary source of the arachidonate used for eicosanoid synthesis in animals, and it is known to be the anchor that can link a variety of proteins to the external leaflet of the plasma membrane via a glycosyl bridge (glycosyl-phosphatidylinositol(GPI)-anchored proteins) . Phosphonolipids are lipids with a phosphonic acid moiety esterified to glycerol, i.e. with a carbonphosphorus bond that is not easily hydrolysed by chemical reagents. Phosphonylethanolamine, for example, is found mainly in marine invertebrates and in protozoa. A ceramide analogue is often found in the same organisms (see below). (More...).

Ether lipids: Many glycerolipids, but mainly phospholipids, and those of animal and microbial origin especially, contain aliphatic residues linked either by an ether bond or a vinyl ether bond to position 1 of L-glycerol. When a lipid contains a vinyl ether bond, the generic term "plasmalogen" is often used. They can be abundant in the phospholipids of animals and microorganisms, and especially in the phosphatidylethanolamine fraction. In this instance, it has been recommended that they should be termed "plasmanylethanolamine" and "plasmenylethanolamine", respectively. On hydrolysis of glycerolipids containing an alkyl ether bond, 1-alkylglycerols are released that can be isolated for analysis. Similarly, when plasmalogens are hydrolysed under basic conditions, 1-alkenylglycerols are released. Aldehydes are formed on acidic hydrolysis. With both groups of compound, the aliphatic residues generally have a chainlength of 16 or 18, and they are saturated or may contain one additional double bond, that is remote from the ether linkage (More...).

'Platelet-activating factor' or 1-alkyl-2-acetylsn-glycerophosphorylcholine is an ether-containing phospholipid, which has been studied intensively because it can exert profound biological effects at minute concentrations. For example, it effects aggregation of platelets at concentrations as low as 10-11 M, and it induces a hypertensive response at very low levels. Also, it is a mediator of inflammation and has messenger functions. (More...). 1-Alkyl-2,3-diacyl-sn-glycerols, analogues of triacylglycerols, tend to be present in trace amounts only in animal tissues, but can be major constituents of certain fish oils. Related compounds containing a 1-alk-1'-enyl moiety ('neutral plasmalogens') are occasionally present also. (More...).

Glycoglycerolipids
In plants, especially the photosynthetic tissues, a substantial proportion of the lipids consists of 1,2-diacyl-sn-glycerols joined by a glycosidic linkage at position sn-3 to a carbohydrate moiety. The main components are the mono- and digalactosyldiacylglycerols, but related compounds have been found with up to four galactose units, or in which one or more of these is replaced by glucose moieties. It is clear that these have an important role in photosynthesis, but many of the details have still to be worked out.

In addition, a 6-O-acyl-monogalactosyldiacylglycerol is occasionally a component of plant tissues. See our web pages dealing with plant galactolipids.

A related unique plant glycolipid is sulfoquinovosyldiacylglycerol or the "plant sulfolipid". It contains a sulfonic acid residue linked by a carbon-sulfur bond to the 6-deoxyglucose moiety of a monoglycosyldiacylglycerol and is found exclusively in the chloroplasts. Monogalactosyldiacylglycerols are not solely plant lipids as they have been found in small amounts in brain and nervous tissue in some animal species. A range of complex glyceroglycolipids have also been characterized from intestinal tract and lung tissue. They exist in both diacyl and alkyl acyl forms. Such compounds are destroyed by some of the methods used in the isolation of glycosphingolipids, so they may be more widespread than has been thought. A complex glyco-glycero-sulfolipid, termed seminolipid, of which the main component is 1-O-hexadecyl-2-O-hexadecanoyl-3-O-(3'-sulfo--D-galactopyranosyl)-sn-glycerol, is the principal glycolipid in testis and sperm. See our web pages on animal glycosyldiacylglycerols. A further range of highly complex glycolipids occur in bacteria and other microorganisms, often with mannose as a carbohydrate moiety. These include acylated sugars that do not contain glycerol.

Sphingomyelin and Glycosphingolipids

Sphingolipids consist of long-chain bases, linked by an amide bond to a fatty acid and via the terminal hydroxyl group to complex carbohydrate or phosphorus-containing moieties.

Long-chain bases (sphingoids or sphingoid bases) are the characteristic structural unit of sphingolipids. They are long-chain (12 to 22 carbon atoms) aliphatic amines, containing two or three hydroxyl groups, and often a distinctive trans-double bond in position 4. The commonest or most abundant of these in animal tissues is sphingosine, ((2S,3R,4E)-2-amino-4-octadecen-1,3-diol) (illustrated). More than a hundred long-chain bases have been found in animals, plants and microorganisms, and many of these may occur in a single tissue, but almost always as part of a complex lipid as opposed to in the free form. The aliphatic chains can be saturated, monounsaturated and diunsaturated, with double bonds of either the cis or trans configuration, and they may sometimes have methyl substituents.

In addition, saturated and monoenoic straight- and branched-chain trihydroxy bases are found. For example, phytosphingosine ((2S,3S,4R)2-amino-octadecanetriol) is the most common long-chain base of plant origin. For shorthand purposes, a nomenclature similar to that for fatty acids can be used, i.e. the chain-length and number of double bonds are denoted in the same manner with the prefix "d" or "t" to designate di- and trihydroxy bases respectively. Thus, sphingosine is d18:1 and phytosphingosine is t18:0. (More...).

Ceramides contain fatty acids linked by an amide bond to the amine group of a long-chain base. In general, they are present at low levels only in tissues, but they are key intermediates in the biosynthesis of the complex sphingolipids. In addition, they have important functions in cellular signalling, and especially in the regulation of apoptosis, and cell differentiation, transformation and proliferation. Unusual ceramides have been located in the epidermis of the pig and humans; the fatty acids linked to the sphingoid base consist of C30 and C32 (-hydroxylated components, with predominantly the essential fatty acid, linoleic acid, esterified to the terminal hydroxyl group. They are believed to have a special role in preventing the loss of moisture through the skin. (More...). Sphingomyelin is a sphingophospholipid and consists of a ceramide unit linked at position 1 to phosphorylcholine; it is found as a major component of the complex lipids of all animal tissues but not of plants or micro-organisms.

It resembles phosphatidylcholine in many of its physical properties, and can apparently substitute in part for this in membranes although it also has its own unique role. For example, it is a major constituent of the plasma membrane of cells, where it is concentrated together with sphingoglycolipids and cholesterol in tightly organized subdomains termed 'rafts'. Sphingosine tends to be the most abundant long-chain base constituent, and it is usually accompanied by sphinganine and C20 homologues.

Sphingomyelin is a precursor for a number of sphingolipid metabolites that have important functions in cellular signalling, including sphingosine-1-phosphate (see below), as part of the 'sphingomyelin cycle'. A correct balance between the various metabolites is vital for good health. Niemann-Pick disease is a rare lipid storage disorder that results from of a deficiency in the enzyme responsible for the degradation of sphingomyelin. (More...).

Ceramide phosphorylethanolamine is found in the lipids of insects and some fresh water invertebrates; the phosphonolipid analogue, ceramide 2-aminoethylphosphonic acid, has been detected in sea anemones and protozoa. Ceramide phosphorylinositol is also found in some organisms, and like phosphatidylinositol, it can be an anchor unit for oligosaccharide-linked proteins in membranes. (More...). Neutral glycosylceramides: The most widespread glycosphingolipids are the monoglycosylceramides (or cerebrosides), and they consist of a basic ceramide unit linked by a glycosidic bond at carbon 1 of the long-chain base to glucose or galactose. They were first found in brain lipids, where the principal form is galactosylceramide, but they are now known to be ubiquitous constituents of animal tissues. Glucosylceramide is also found in animal tissues, and especially in skin, where it functions as part of the water permeability barrier. It is the biosynthetic precursor of lactosylceramide, and thence of the complex oligoglycolipids and gangliosides. In addition, glucosylceramide is found in plants, where the main long-chain base is phytosphingosine.

O-Acyl-glycosylceramides have been detected in small amounts in some tissues, as have cerebrosides with monosaccharides such as xylose, mannose and fucose. (More...). Di-, tri- and tetraglycosylceramides (oligoglycosylceramides) are present in most animal tissues at low levels. The most common diglycosyl form is lactosylceramide, and it can be accompanied by related compounds containing further galactose or galactosamine residues. Tri- and tetraglycosylceramides with a terminal galactosamine residue are sometimes termed "globosides", while glycolipids containing fucose are known as "fucolipids". Lactosylceramide is the biosynthetic precursor of most of these with further monosaccharide residues being added to the end of the carbohydrate chain (up to as many as twenty). They are an important element of the immune response system. For example

some glycolipids are involved in the antigenicity of blood group determinants, while others bind to specific toxins or bacteria. As the complex glycosyl moiety is considered to be of primary importance in this respect, it has received most attention from investigators. However, certain of these lipids have been found on occasion to have distinctive long-chain base and fatty acid compositions, which enhance their biological activity. Some glycolipids accumulate in persons suffering from rare disease syndromes, characterized by deficiencies in specific enzyme systems related to glycolipid metabolism. Sulfate esters of galactosylceramide and lactosylceramide (sulfoglycosphingolipids often referred to as "sulfatides" or "lipid sulfates"), with the sulfate group linked to position 3 of the galactosyl moiety, are major components of brain lipids and they are found in trace amounts in other tissues. (More...). Complex plant sphingolipids, phytoglycosphingolipids, containing glucosamine, glucuronic acid and mannose linked to the ceramide via phosphorylinositol, were isolated and characterized from seeds initially, but related compounds are also known to be present in other plant tissues and in fungi. Gangliosides are highly complex oligoglycosylceramides, which contain one or more sialic acid groups (N-acyl, especially acetyl, derivatives of neuraminic acid, abbreviated to "NANA") in addition to glucose, galactose and galactosamine.

The polar and ionic nature of these lipids renders them soluble in water (contrary to some definitions of a lipid). They were first found in the ganglion cells of the central nervous system, hence the name, but are now known to be present in most animal tissues. The long-chain base and fatty acid components of gangliosides can vary markedly between tissues and species, and they are presumably related in some way to function. Gangliosides have been shown to control growth and differentiation of cells, and they have important roles in the immune defence systems. They act as receptors for a number of tissue metabolites and in this way may regulate cell signalling. Also, they bind specifically to various bacterial toxins, such as those from botulinum, tetanus and cholera. A number of unpleasant lipidoses have been identified involving storage of excessive amounts of gangliosides in tissues, the most important of which is Tay-Sachs disease. (More...). Sphingosine-1-phosphate is one of the simplest sphingolipids structurally. It is present at low levels only in animal tissues, but it is a pivotal lipid in many cellular signalling pathways (together with ceramide and ceramide-1-phosphate). For example, within cells,

sphingosine-1-phosphate promotes cellular division (mitosis), while in the blood it may play a critical role in platelet aggregation and thrombosis. (More...).

The fatty acids of sphingolipids: Although structures of fatty acids are discussed in greater depth below, it is worth noting that the acyl groups of ceramides are very different from those in the glycerolipids. They tend to consist of long-chain (C16 up to C26 but occasionally longer) odd- and even-numbered saturated or monoenoic fatty acids and related 2-D-hydroxy fatty acids, both in plant and animal tissues. Linoleic acid may be present at low levels in sphingolipids from animal tissues, but polyunsaturated compounds are rarely found (although their presence is often reported in error).

Fatty Acids
The common fatty acids of plant tissues are C16 and C18 straight-chain compounds with zero to three double bonds of a cis (or Z) configuration. Such fatty acids are also abundant in animal tissues, together with other even numbered components with a somewhat wider range of chain-lengths and up to six cis double bonds separated by methylene groups (methylene-interrupted). The systematic and trivial names of those fatty acids encountered most often, together with their shorthand designations, are listed in the table.
The common fatty acids of animal and plant origin

Systematic name Saturated fatty acids ethanoic butanoic hexanoic octanoic decanoic dodecanoic tetradecanoic hexadecanoic octadecanoic eicosanoic docosanoic Monoenoic fatty acids cis-9-hexadecenoic

Trivial name Shorthand

acetic butyric caproic caprylic capric lauric myristic palmitic stearic arachidic behenic palmitoleic

2:0 4:0 6:0 8:0 10:0 12:0 14:0 16:0 18:0 20:0 22:0 16:1(n-7)

cis-6-octadecenoic petroselinic cis-9-octadecenoic oleic cis-11-octadecenoic cis-vaccenic cis-13-docosenoic erucic cis-15-tetracosenoic nervonic Polyunsaturated fatty acids* 9,12-octadecadienoic linoleic 6,9,12-octadecatrienoic -linolenic 9,12,15-octadecatrienoic -linolenic 5,8,11,14-eicosatetraenoic arachidonic 5,8,11,14,17-eicosapentaenoic EPA 4,7,10,13,16,19-docosahexaenoic DHA * all the double bonds are of the cis configuration

18:1(n-12) 18:1(n-9) 18:1(n-7) 22:1(n-9) 24:1(n-9) 18:2(n-6) 18:3(n-6) 18:3(n-3) 20:4(n-6) 20:5(n-3) 22:6(n-3)

The most abundant saturated fatty acid in nature is hexadecanoic or palmitic acid. It can also be designated a "16:0" fatty acid, the first numerals denoting the number of carbon atoms in the aliphatic chain and the second, after the colon, denoting the number of double bonds. All the even-numbered saturated fatty acids from C2 to C30 have been found in nature, but only the C14 to C18 homologues are likely to be encountered in appreciable concentrations in glycerolipids, other than in a restricted range of commercial fats and oils. Oleic or cis-9-octadecenoic acid, the most abundant monoenoic fatty acid in nature, is designated as "18:1", or more precisely as 9c-18:1 or as 18:1(n-9) (to indicate that the last double bond is 9 carbon atoms from the terminal methyl group).

The latter form of the nomenclature is of special value to biochemists. Similarly, the most abundant cis monoenoic acids fall into the same range of chain-lengths, i.e. 16:1(n-7) and 18:1(n-9), though 20:1 and 22:1 are abundant in fish. Fatty acids with double bonds of the trans (or E) configuration are found occasionally in natural lipids, or are formed during food processing (hydrogenation) and so enter the food chain, but they tend to be minor components only of animal tissue lipids, other than of ruminants, where they are formed naturally by biohydrogenation. Their suitability for human nutrition is currently a controversial subject. The C18 polyunsaturated fatty acids, linoleic or cis-9,cis-12-octadecadienoic acid (18:2(n6)) and -linolenic or cis-9,cis-12,cis-15-octadecatrienoic acid (18:3(n-3)), are major

components of most plant lipids, including many of the commercially important vegetable oils.

They are essential fatty acids in that they cannot be synthesised in animal tissues. On the other hand, as linoleic acid is almost always present in foods, it tends to be relatively abundant in animal tissues. In turn, these fatty acids are the biosynthetic precursors in animal systems of C20 and C22 polyunsaturated fatty acids, with three to six double bonds, via sequential desaturation and chain-elongation steps (desaturases in animal tissues can only insert a double bond on the carboxyl side of an existing double bond). Those fatty acids derived from linoleic acid, especially arachidonic acid (20:4(n-6)), are important constituents of the membrane phospholipids in mammalian tissues, and are also the precursors of the prostaglandins and other eicosanoids. In fish, linolenic acid is the more important essential fatty acid, and polyunsaturated fatty acids of the (n-3) series, especially eicosapentaenoic acid (20:5(n-3) or EPA) and docosahexaenoic acid (22:6(n-3) or DHA), are found in greater abundance.

Many other fatty acids that are important for nutrition and health do of course exist in nature, and at present there is great interest in -linolenic acid (18:3(n-6)), available from evening primrose oil -

- and in conjugated linoleic acid (mainly 9-cis,11-trans-octadecadienoate) or 'CLA', a natural constituent of dairy products, that is claimed to have remarkable health-giving properties.

Branched-chain fatty acids are synthesised by many microorganisms (most often with an iso- or an anteiso-methyl branch) and they are synthesised to a limited extent in higher organisms. They enter animal tissues via the diet, especially with ruminants.

Phytanic acid, 3,7,11,15-tetramethylhexadecanoic acid, is a metabolite of phytol and is found in animal tissues, but generally at low levels only. Fatty acids with many other substituent groups are found in certain plants and microorganisms, and they may be encountered in animal tissues, which they enter via the food chain. These substituents include acetylenic and conjugated double bonds, allenic groups, cyclopropane, cyclopropene, cyclopentene and furan rings, and hydroxy-, epoxyand keto-groups. For example, 2-hydroxy fatty acids are synthesised in animal and plant tissues, and are often major constituents of the sphingolipids. 12-Hydroxy-octadec-9enoic or 'ricinoleic' acid is the main constituent of castor oil.

Eicosanoids and Related Lipids

The term eicosanoid is used to embrace biologically active lipid mediators (C20 fatty acids and their metabolites), including prostaglandins, thromboxanes, leukotrienes and other oxygenated derivatives, which exert their effects at very low concentrations. They are produced primarily by three classes of enzymes, cyclooxygenases (COX-1 and COX-2), lipoxygenases (LOX) and cytochrome P450 epoxygenase. The key precursor fatty acids are 8c,11c,14c-eicosatrienoic (dihomo--linolenic or 20:3(n-6)), 5c,8c,11c,14ceicosatetraenoic (arachidonic or 20:4(n-6)) and 5c,8c,11c,14c,17c-eicosapentaenoic (20:5(n-3) or EPA) acids (see our web page on 'polyunsaturated fatty acids'). More recently docosanoids (resolvins and protectins) derived from 4c,7c,10c,13c,16c,19cdocosahexaenoic acid (22:6(n-3) or DHA) have been described. Other eicosanoids are produced by non-enzymic means (isoprostanes).

Those derived from arachidonic acid appear to be of special importance and have been most studied. The prostaglandins and thromboxanes have cyclic structures, generated by cyclo-oxygenase enzymes, and are involved in the processes of inflammation. The hydroxy-eicosatetraenoic acids are generated by lipoxygenases, and of these the 5lipoxygenase is especially important as it produces the first intermediate in the biosynthesis of leukotrienes. The resolvins and protectins have anti-inflammatory properties.

Plant products, such as the jasmonates and other oxylipins derived from 9c,12c,15c-octadecatrienoic (-linolenic or 18:3(n-3)) acid are also generated by the action of lipoxygenases. They are involved in responses to physical damage by animals or insects, stress and attack by pathogens. There are obvious structural similarities between the jasmonates and prostanoids. Our introductory page on eicosanoids will lead you to further information. Of course, many more lipids occur in nature than can be described in this document. I have not touched on proteolipids, lipoproteins and lipopolysaccharides here, for example, but there is information on these and other lipids elsewhere on this website. New lipids continue to be found, and no doubt many remain to be discovered. Other than the Lipid Library, the book "Les Lipides dans le Monde Vivant. Introduction la Lipidomique" by Claude Leray (Lavoisier, France, 2010) and the website http://www.cyberlipid.org are the best sources of information on lipid structures.

WHAT LIPIDS DO
Their Biological Functions
For many years, lipids were considered to be intractable and uninteresting oily materials with two main functions to serve as a source of energy and as the building blocks of membranes. They were certainly not considered to be appropriate candidates for such important molecular tasks as intracellular signalling or local hormonal regulation. In 1929, George and Mildred Burr demonstrated that linoleic acid was an essential dietary constituent, but it was many years before the importance of this finding was recognized by biochemists in general. With the discovery by Bergstrm, Samuelsson and others in 1964 that the essential fatty acid arachidonate was the biosynthetic precursor of the prostaglandins with their effects on inflammation and other disease states, the scientific world in general began to realize that lipids were much more interesting than they had previously thought.

A major milestone was achieved in 1979 with the discovery of the first biologically active phospholipid, platelet-activating factor. At about the same time, there arose an awareness of the distinctive functions of phosphatidylinositol and its metabolites. Since then, virtually every individual lipid class has been found to have some unique biological role that is distinct from its function as a source of energy or as a simple construction unit of a membrane. Indeed it is now recognised that lipids in membranes function also in the trafficking of cellular constituents, the regulation of the activities of membrane proteins and signalling. All multi-cellular organisms, use chemical messengers to send information between organelles and to other cells and as relatively small hydrophobic molecules, lipids are excellent candidates for signalling purposes. The fatty acid constituents have welldefined structural features, such as cis-double bonds in particular positions, which can carry information by binding selectively to specific receptors. In esterified form, they can infiltrate membranes or be translocated across them to carry signals to other cells. During transport, they are usually bound to proteins so their effective solution concentrations are very low, and they are can be considered to be inactive until they reach the site of action and encounter the appropriate receptor.

Storage lipids, such as triacylglycerols, in their cellular context are inert, and indeed esterification with fatty acids may be a method of de-activating steroidal hormones, for example, until they are actually required. In contrast, polar phospholipids have both hydrophobic and hydrophilic sites that can bind via various mechanisms to membrane proteins and influence their activities. Glycosphingolipids carry complex carbohydrate moieties that have a part to play in the immune system, for example. Lipids have been implicated in a number of human disease states, including cancer and cardiovascular disease, sometimes in a detrimental and sometimes in a beneficial manner. In short, every scientist should now be aware that lipids are just as fascinating as all the other groups of organic compound that make up living systems. In this web document, the main biological functions of some key lipids are briefly summarized to give a general overview, but much more information is available here on those pages dealing with specific lipid classes.

Fatty Acids
Fatty acids are one of the defining constituents of lipids and are in large part responsible for the distinctive physical and metabolic properties of the latter. However, they are also important in non-esterified form, i.e. as free (unesterified) fatty acids. They are released from triacylglycerols during fasting to provide a source of energy and of structural components for cells (see below), where they are of course of vital importance. However, it has become evident that there are a number of more dynamic functions of fatty acids, which are attracting great interest. It has long been known that linoleic and linolenic acids are essential fatty acids, in that they cannot be synthesised by animals and must come from plants via the diet. They are precursors of arachidonic, eicosapentaenoic and docosahexaenoic acids, which are vital components of all membrane lipids. However, we cannot even now claim to fully understand the reasons for the unique requirements for these fatty acids. Dietary fatty acids of short and medium chain-length are not usually esterified but are oxidized rapidly in tissues as a source of 'fuel' to support all the events necessary to keep organisms functioning. Longer-chain fatty acids are usually esterified first to triacylglycerols or structural lipids in tissues. Although all lipids are in a state of dynamic flux, membrane lipids are conserved in content and composition in essence, except under conditions of extreme stress. Triacylglycerols are the primary storage form of long-chain fatty acids for energy and structural purposes, and free acids can be mobilized quickly when required for transport in an appropriate form to the heart, liver and other tissues where they can be oxidized.

Polyunsaturated fatty acids are important as constituents of the phospholipids, where they appear to confer distinctive properties to the membranes, in particular by decreasing their rigidity. The presence of saturated and monoenoic acids ensure that there is a correct balance between rigidity and flexibility. Indeed, saturated and 2-hydroxy fatty acids in sphingolipids appear to give additional rigidity and hydrogen-bonding stability to the sub-domains of membranes termed 'rafts'.

The essential fatty acids, linoleic and linolenic acids and their longer-chain polyunsaturated metabolites, such as arachidonic acid, can be found in most lipid classes, but they are also the precursors of many different types of eicosanoids, including the hydroxyeicosatetraenes, prostanoids (prostaglandins, thromboxanes and prostacyclins), leukotrienes (and lipoxins) and resolvins, not to forget the isoprostanes, which are formed by non-enzymic means. It is surely no coincidence that plant hormones, such as the jasmonates, are also derived from the essential fatty acids and have structural similarities. While, they are usually treated separately in biochemical textbooks, it should not be forgotten that these compounds are in fact fatty acids. Some of them are occasionally found esterified to phospholipids (and glycosyldiacylglycerols in plants), although their short half-lives may preclude long-term storage in this form. For example, the isoprostanes are all formed in situ in lipids within membranes. The eicosanoids are highly potent at nanomolar concentrations in the regulation of innumerable biological activities, especially in relation to inflammatory responses, pain and fever. Fatty acids are also the biosynthetic precursors of many insect pheromones and of secondary metabolites in plants. Unesterified fatty acids can act as second messengers required for the translation of external cellular signals, as they are produced rapidly as a consequence of the binding of specific agonists to plasma membrane receptors. Within cells, fatty acids can act to amplify or otherwise modify signals to influence the activities of such enzymes as protein kinases, phospholipases, and many more. They are involved in regulating gene expression, mainly targeting genes that encode proteins with roles in fatty acid transport or metabolism via effects on transcription factors, i.e. peroxisome proliferator-activated receptors (PPARs) in the nuclei of cells. Such effects can be highly specific to particular fatty acids. Thus, unesterified arachidonic acid may have some biological importance per se as part of the mechanism by which apoptosis (programmed cell death) is regulated.

Tri-, Di- and Monoacylglycerols

Virtually all the natural fats and oils of commerce consist of triacylglycerols, but here we are concerned with their biological functions. As discussed briefly above triacylglycerols are the storage lipid in animal and plant cells, where they occur as discrete droplets surrounded by a protective monolayer of phospholipids and functional hydrophobic proteins. Fatty acids are released when required by hydrolysis reactions catalysed by lipases under the influence of hormones, though a high proportion is usually re-esterified to biochemically inert triacylglycerols for extracellular transport. One specialized form of adipose tissue, brown fat, is highly vascularized and rich in mitochondria, which oxidize fat so rapidly that heat is generated. This appears to be especially important in young animals and in those recovering from hibernation. Triacylglycerols are the main lipid component in the only material designed by nature entirely as a food, i.e. milk, though triacylglycerols in seeds could perhaps be considered similarly as 'food' for the developing plant embryo until it is capable of photosynthesis.

However, triacylglycerol depots have other functions. Subcutaneous depots serve as insulation against cold in many terrestrial animals, as is obvious in the pig, which is surrounded by a layer of fat, and it is especially true for marine mammals. In the latter and in fish, the lipid depots are less dense than water and so aid buoyancy with the result that less energy is expended in swimming. More surprisingly, perhaps, triacylglycerols together with the structurally related glyceryl ether diesters and wax esters are the main components of the sonar lens used in echolocation by dolphins and some whales. sn-1,2-Diacylglycerols are formed as intermediates in the biosynthesis of triacyl-snglycerols and via the action of a diacylglycerol kinase of phospholipids. In addition, they function as second messengers in many cellular processes, modulating vital biochemical mechanisms by activating members of the protein kinase C family of enzymes. They are formed together with the important inositol phosphates by the action of the enzyme phospholipase C on phosphatidylinositol and polyphosphoinositides mainly. Their influence is believed to extend to the pathophysiology of cancer and other disease states. 2-Monoacylglycerols are produced when triacylglycerols are digested in the intestines of animals, but they are re-esterified before they are transported elsewhere in the body. In

general, monoacylglycerols are minor components of tissues, which are never permitted to accumulate because their strong detergent properties would have a disruptive effect on membranes.

2-Arachidonoylglycerol, a further product of phosphatidylinositol catabolism, is important in animal tissues as an endogenous ligand for cannabinoid receptors and as a mediator of the inflammatory response.

Waxes
Waxes form a thin layer over all the green tissue of plants that is both a chemical and a physical barrier. This layer serves many purposes, for example to limit the diffusion of water and solutes, while permitting a controlled release of volatiles that may deter pests or attract pollinating insects. It provides protection from disease and insects, and helps the plants resist drought. Waxes also have a waterproofing and protective role for insects.

Waxes can have a storage function, as in marine organisms and for example in the seeds of the jojoba plant. Bees use wax to produce the rigid structures of honeycombs. The uropygial glands of birds secrete waxes, which they use to provide water-proofing for feathers.

Some Other Simple Lipids

Before a fatty acid can be metabolized in tissues, it must usually be activated by conversion to a Coenzyme A ester or acyl-CoA, with the fatty acid group linked to the terminal thiol moiety. The thiol ester is a highly energetic bond that permits a facile transfer of the acyl group to receptor molecules. Acyl-carnitines assist the transport and metabolism of fatty acids in and out of mitochondria, where they are oxidized for energy production. In so doing, carnitine maintains a balance between free and esterified coenzyme A in cells.

Long-chain N-acylethanolamines are ubiquitous trace constituents of animal and human cells with important pharmacological properties. The nature of the fatty acid controls the biological functions. Anandamide or Narachidonoylethanolamine has attracted special interest, because of its marked biological activities, exerting its effects through binding to and activating specific cannabinoid receptors. Like 2-arachidonoylglycerol, discussed above, it is an endogenous cannabinoid or endocannabinoid. In contrast, oleoylethanolamide is an endogenous regulator of food intake with potential as an anti-obesity drug, while palmitoylethanolamide is an antiinflammatory agent, and stearoylethanolamide is a pro-apoptotic agent. Similarly, changing the nature of the amide moiety also changes the function. Thus, the simple oleamide molecule or cis-9,10-octadecenamide, isolated from the cerebrospinal fluid of sleep-deprived cats, has been identified as the signalling molecule responsible for causing sleep. Many simple fatty acid derivatives of amino acids are now known and their biological functions are slowly being revealed.

Sterols

Cholesterol is a ubiquitous component of all animal tissues, where much of it is located in the membranes. It occurs in the free form and esterified to long-chain fatty acids (cholesterol esters) in animal tissues, including the plasma lipoproteins. It is generally believed that the main function of cholesterol is to modulate the fluidity of membranes by interacting with their complex lipid components, specifically the phospholipids such as phosphatidylcholine and sphingomyelin, increasing the degree of order by promoting a 'liquid-ordered phase'. There is also considerable evidence for more intimate protein-cholesterol interactions that may regulate the activities of certain membrane proteins. It is of course the precursor of bile acids, vitamin D and steroidal hormones.. In plants, cholesterol tends to be a minor component only of a complex phytosterol fraction that includes campesterol, -sitosterol, stigmasterol, 5-avenasterol and brassicasterols, while yeasts and fungi tend to contain ergosterol as their main sterol component. Plant sterols are also able to regulate membrane fluidity and permeability,

and they modulate the activity of membrane-bound enzymes. Some bacterial species produce structurally and functionally related lipids, the hopanoids.

Complex Lipids in Membranes

Cellular membranes are semi-permeable barriers that enclose and define the cell and its organelles. They control the transport of materials, including signalling molecules, and indeed many reactions occur within membranes, including energy production and biosynthesis of cellular components. In addition, they can deform to enable budding, fission and fusion. It is evident that the specific lipid compositions of membranes have evolved to provide a barrier to the diffusion of ionic solutes and other molecules into cellular compartments where they may not be required. At the same time, the membrane environment provides a stable molecular platform for essential metabolic events and for intense signalling activity. Cellular membranes are the first site for receipt of extracellular signals, they recruit and activate effector molecules, and they are the launch pad for activated effector molecules throughout the cell. The characteristic feature of membrane lipids, which are essential for all of these functions, is that they contain both hydrophobic and hydrophilic constituents, i.e. they are amphiphilic. As such, they are weak surfactants and they tend to form aggregates in bilayer or hexagonal-II arrangements in aqueous media in the normal temperature ranges that prevail in living cells. However, in natural membranes, there is a mixture of lipid types, which determine that bilayer structures predominate.

Glycerophospholipids, such as phosphatidylcholine, phosphatidylethanolamine and so forth, together with the sphingolipids, such as sphingomyelin and the glycosphingolipids, and cholesterol are essential structural elements of all biological membranes. In the conventional model, which is illustrated here in two dimensions, polar lipids form a bilayer with the polar head groups oriented towards the aqueous phase while the hydrophobic fatty acyl moieties are arranged internally.

Proteins, such as enzymes, transport systems or signalling receptors, can span the bilayer and take up a considerable proportion of the membrane surface. They interact via their basic amino acid residues with the ionic groups of polar lipids via electrostatic interactions, generating a net charge that is mainly negative or zwitterionic. In the process, their biological properties may be modified. These membrane structures are not static, and free movement is possible within each leaflet (lateral diffusion) and between leaflets (vertical or flip-flop diffusion). In addition, lipid molecules can rotate around their principal axis (rotational diffusion). The lateral and rotational diffusions are responsible for the liquid characteristics of membranes, with the constraint that the hydrophobic chains remain parallel to each other and perpendicular to the surface of the bilayer. The distribution of lipids in each of the membrane leaflets is asymmetric with phosphatidylcholine and sphingolipids located in the outer leaflet of the plasma membrane, for example, while phosphatidylethanolamine and anionic phospholipids such as phosphatidylinositol (and polyphosphoinositides) and phosphatidylserine occur primarily in the inner leaflet. Cholesterol occurs in roughly equal proportions in both faces, where it modulates the fluidity of membranes by its interaction with phospholipids. A membrane translocation machinery, which consumes large amounts of energy, is required to maintain this asymmetry. Each glycerophospholipid with its distinctive polar head group and characteristic fatty acid composition modifies the properties of a membrane in a unique manner and contributes to its overall properties. Phosphatidylcholine is often the most abundant lipid in membranes, and it has a cylindrical shape, which does not induce curvature. On the other hand, an increased concentration of cone-shaped lipids on one side and inverted cones on the other side of a membrane will bring about curvature, which may be required for membrane transport and fusion processes.

The balance between saturated, monoenoic and polyunsaturated fatty acids is important in maintaining the optimum degree of fluidity of a given membrane. Docosahexaenoic acid, for example, adopts a more flexible and compact conformation than more saturated chains with an average length only 60% of that for oleic chains, and this in turn increases the conformational disorder of saturated chains in mixed-chain phospholipids. In bacterial membranes, branched-chain and cyclopropane fatty acids modify the fluidity in an analogous manner. When ether and plasmalogen forms of lipids are also taken into account, membranes can contain a thousand distinct molecular species of phospholipid. It is obviously impossible to quantify the relative importance of each of

these to the physical and biological properties of membranes, and some general assessments only are possible. While the need to form stable bilayers is a primary prerequisite for all membranes, there is also a requirement for a potential ability of the lipids to form non-bilayer structures for some membrane-associated cell processes. For example, short-lived non-bilayer structures with specific lipid components are probably formed in the processes of fusion and fission of lipid bilayers and for cell division, while the activities of certain membrane-associated proteins can be modulated by lipids that do not form lamellar layers. Further complexity is imposed by specific associations of phospholipids with membrane proteins. Defined lipid species are required to stabilize protein structures, to control the insertion and folding of proteins and membranes, and even for the assembly or polymerization of enzyme complexes with direct effects on their functions. In addition, many proteins are directed to membranes by covalent linkages to lipids, such as the glycosyl phosphatidylinositol anchors, or by modification with myristoyl, palmitoyl, prenyl or sterol moieties (see below). The sphingolipids together with cholesterol arrange themselves into distinct sub-domains or 'rafts' (see below) with certain membrane enzymes, and they act to compartmentalize these and their different biochemical functions of course.

Glycerophospholipids
Phospholipids play multiple roles in cells other than by establishing permeability barriers. For example, they provide a matrix for the assembly and function of a wide variety of enzymes, they participate in the synthesis of macromolecules, and they act as molecular signals to influence metabolic events. Anionic lipids like phosphatidylinositol and its phosphorylated derivatives, which are concentrated on the cytoplasmic leaflet of membranes, exert a control on the properties of the membranecytosol interface and consequently on many aspects of membrane trafficking, including vacuole formation transport and fusion. Specific lipids of this kind are associated with particular organelles, where in combination with other signalling molecules they can recruit effector proteins with appropriate functions for each cellular compartment. Phosphatidylcholine is a zwitterionic lipid and usually the most abundant phospholipid in membranes of animal and plants, constituting a high proportion of the outer leaflet of the plasma membrane. It is also an integral component of the lipoproteins in plasma. However, it may serve as a source of diacylglycerols with a signalling function, while the plasmalogen form especially may provide arachidonate for eicosanoid production. In addition, phosphatidylcholine is the biosynthetic precursor of sphingomyelin (see below) and many other signalling molecules and thus has an influence on innumerable metabolic pathways.

Platelet-activating factor or 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine, a closely related lipid, was the first biologically active phospholipid to be discovered. Amongst the innumerable activities that have been documented, it effects the aggregation of platelets at concentrations as low as 10-11 M, it is a mediator of inflammation and it is involved in the mechanism of the immune response. Phosphatidylethanolamine is also a major component of membranes, especially in bacteria, with distinctive physical properties because of its small head group and hydrogen bonding capacity. In the bacterium E. coli, it supports active transport by the lactose permease, and other transport systems may require or be stimulated by it. In animal and plants, it acts as a 'chaperone' during the assembly of membrane proteins to guide the folding path for the proteins and to aid in the transition from the cytoplasmic to the membrane environment. Phosphatidylinositol is an acidic or anionic phospholipid, a high proportion of which in animal membranes consists of the 1-stearoyl,2-arachidonoyl molecular species, which is of considerable biological importance. It is the primary donor of 1,2-diacylglycerols with their specific signalling functions (see above), and of inositol phosphates with many different biological activities. In addition, it is the main source of arachidonic acid for the production of eicosanoids and of endogenous cannabinoids. In all eukaryotes, phosphatidylinositol serves as an anchor that links a variety of proteins to the external leaflet of the plasma membrane via complex glycosyl bridges, i.e. glycosylphosphatidylinositol(GPI)-anchored proteins. A further acidic lipid, phosphatidylserine, contributes substantially to non-specific electrostatic interactions in the inner leaflet of membranes. This normal distribution is disturbed during platelet activation and in the process of cellular apoptosis when the lipid is transferred from the inner to the outer leaflet of the plasma membrane and acts as a signal to scavenger cells. Phosphatidylserine chelates with calcium to act as the foundation for bone growth. It is also an essential cofactor for the activation of many enzymes, including protein kinase C, which is a key enzyme in signal transduction.

Cardiolipin or diphosphatidylglycerol is a unique acidic phospholipid with four acyl groups. In the mitochondria of cells, its primary location, many biological functions of this lipid have been identified, but the main ones involve activation of those enzymes concerned with oxidative phosphorylation. Indeed, it is integrated into their quaternary structure, where it is an essential component of the interface between the enzymes and their environment and may stabilize the active sites. In higher plants, cardiolipin is an integral constituent of the photosystem II complexes, which are also involved in oxidative processes, and where it may be required for the maintenance of structural and functional properties. Phosphatidic acid is generally a minor component of cells, but it is a key intermediate in the biosynthesis of all other phospholipids. It is known to have signalling functions in animal cells, by specific binding to particular proteins, and it may be even more important in higher plants where it is formed rapidly in response to stresses of all kinds. Lysophospholipids, i.e. with only one mole of fatty acid per mole of lipid, were long thought to be merely intermediates in the biosynthesis of phospholipids that were potentially disruptive to cells if allowed to accumulate, because of their powerful detergent properties. However, lysophosphatidic acid has been shown to have signalling and other biological effects that are dependent on receptor mechanisms. It is produced by a wide variety of cell types and most mammalian cells express receptors for it. For example, it is involved in the activation of protein kinases, adenyl cyclase and phospholipase C, in the release of arachidonic acid, and much more. Interest was stimulated especially by a finding that lysophosphatidic acid is significantly elevated in the plasma of ovarian cancer patients compared to healthy controls, so that it may represent a useful marker for the early detection of the disease. Other lysophospholipids including the sphingolipid analogue, sphingosine-1-phosphate (see below), exhibit a related range of activities. Lysobisphosphatidic acid has a unique stereochemistry and distinctive biological functions.

Glycosyldiacylglycerols
Mono- and digalactosyldiacylglycerols, and sulfoquinovosyldiacylglycerol are important components of membranes of chloroplasts and related organelles, and indeed these are the most abundant lipids in all photosynthetic tissues, including those of higher plants, algae and certain bacteria. They may substitute in part for phospholipids, especially when

phosphorus is limiting, although the distinctive ability of monogalactosyldiacylglycerols to form inverted micelles may be important for membrane structure and for interactions with specific proteins. The thylakoid membrane where photosynthesis occurs in plants has an asymmetric distribution of glycolipids, with much of the digalactosyldiacylglycerol on the luminal leaflet, where it may assist the movement of protons along the membrane surface to the ATPase. While many different functions have been ascribed to these lipids, it is clear that their primary importance is in their interactions with the photosynthetic apparatus.

Glycosyldiacylglycerols have also been found in animal tissues, though usually in rather small amounts, and their role in mammalian membranes is poorly understood. However, seminolipid or 1-O-hexadecyl-2-O-hexadecanoyl-3-O--D-(3'-sulfo)-galactopyranosylsn-glycerol, which was first found in mammalian spermatozoa and testes, is known to be essential for spermatogenesis and may have a role in myelination.

Sphingolipids
Sphingolipids are distinguished by the presence of a long-chain or sphingoid base, such as sphingosine, to which a fatty acid is linked by an amide bond, and usually with the primary hydroxyl group attached to complex phosphoryl or carbohydrate moieties. They have an immense range of functions in tissues that are quite distinct from those of the complex glycerolipids. For example, sphingomyelin has structural similarities to phosphatidylcholine, but has very different physical and biological properties, while the complex oligoglycosylceramides and gangliosides have no true parallels among the glycerolipids.

Free sphingoid bases are found trace levels only in tissues, but they are mediators of a number of cellular events. For example, they inhibit the enzyme protein kinase C, and they are inhibitors of cell growth, although they stimulate cell proliferation and DNA synthesis. Some of the structural features of the long-chain bases are only introduced

after they are esterified with long-chain fatty acids to form ceramides, which are also the primary precursors of the complex sphingolipids. In addition, ceramides have an important role in cellular signalling, and especially in the regulation of apoptosis, and cell differentiation, transformation and proliferation. In contrast, sphingosine-1-phosphate is an especially important sphingosine metabolite and promotes cellular division (mitosis) as opposed to apoptosis, so that the balance between the former and ceramide, ceramide1-phosphate and sphingosine levels in cells is critical. In fact, the biosynthesis and catabolism of sphingolipids involves a large number of metabolites, many of which have distinctive biological activities. In animals the relationships between these metabolites have been rationalized in terms of a 'sphingomyelin cycle', in which each of the various compounds has characteristic metabolic properties. Similar pathways occur in plants although sphingomyelin is not involved. Sphingomyelin is by far the most abundant sphingolipid in animal tissues. In addition to serving as a source of key cellular metabolites, sphingomyelin is an important building block of membranes and like its glycerolipid analogue phosphatidylcholine tends to be most abundant in the plasma membrane of cells and especially in the outer leaflet. The sphingolipids in general contain high proportions of longer-chain saturated and monoenoic fatty acids, often accompanied by high proportions of 2-hydroxy but not polyunsaturated fatty acids.

Sphingomyelin and other sphingolipids together with cholesterol are located in an intimate association in specific sub-domains or 'rafts' (or related structures termed 'caveolae') of membranes. These are laterally segregated regions that form as a result of selective affinities between sphingolipids and membrane proteins. As sphingolipids containing long saturated acyl chains, they pack more tightly together, thus giving sphingolipids much higher melting temperatures than glycerophospholipids. This tight acyl chain packing is essential for raft lipid organization, since the differential packing facility of sphingolipids and cholesterol in comparison with glycerophospholipids leads to phase separation in the membrane, giving rise to the sphingolipid-rich regions ('liquidordered' phase) surrounded by glycerophospholipid-rich domains ('liquid-disordered' phase). The ordered phases are relatively resistant to attack by detergents, a property that is sometimes used to define them. An important result of this process is that rafts contain a variety of different proteins, including glycosyl-phosphatidylinositol(GPI)-anchored proteins and tyrosine receptor kinases. These provide much of the important biological properties of rafts, and are also essential to maintain their stability. Micro-domains or rafts that are enriched in

sphingolipids (other than sphingomyelin), sterols and specific proteins have also been detected in the plasma membrane of plant cells. Monoglycosylceramides or cerebrosides are common constituents of membranes of animals and plants. Galactosylceramide is the principal glycosphingolipid in brain tissue and myelin, while glucosylceramide is a major constituent of skin lipids, and is the source of the unusual complex ceramides that are found in the stratum corneum. It is also the biosynthetic precursor of the oligoglycosphingolipids. Harmful quantities of glucosylceramide accumulate in tissues of patients with Gaucher's disease, an inherited metabolic disorder. In plants, specific glucosylceramides elicit defence responses against fungal attack, and they appear to assist plants to withstand stresses brought about by cold and drought.

The membranes of animals and plants contain a wide range of complex oligoglycosylceramides (several hundred different head groups). Most of these occur on the external leaflet of the plasma membrane in rafts, where they are important components of the body's immune defence system, both as cellular immunogens and as antigens. For example, certain glycolipids are involved in the antigenicity of blood group determinants, while others bind to specific toxins or bacteria. Some function as receptors for cellular recognition, and they can be specific for particular tissues or tumours. Glycosphingolipid sulfates are highly polar acidic molecules that are important in the transport of sodium and potassium ions and osmoregulation in animal tissues; they may also have a role in the protection of the intestinal mucosa against digestive enzymes. Gangliosides are complex oligoglycosylceramides containing sialic acid residues so they are also highly polar and acidic. They are cell-type specific antigens that control the growth and differentiation of cells and have an important role in the interactions between cells, especially in the immune defence systems. They are especially important for myelination in brain and other nervous tissues. In addition, gangliosides act as receptors of interferon, epidermal growth factor, nerve growth factor, insulin and many other metabolites, and in this way they regulate cell signalling. Certain gangliosides bind to specific bacterial toxins and they mediate interactions between microbes and host cells during infections.

Proteolipids and Lipoproteins

Proteins that contain covalently bound fatty acids or other lipid moieties, such as isoprenoids, cholesterol and glycosylphosphatidylinositol, are widespread in nature with many important functions. The term proteolipid is used to define such complexes, and to differentiate them from the plasma lipoproteins. Two main types of protein with a fatty acid modification have been described, i.e. those with only myristoyl and those with predominantly palmitoyl moieties, each with a distinctive type of linkage, amide or thiol ester, respectively. The prenylated lipids contain an isoprenoid group, farnesyl or geranylgeranyl, linked via a sulfur atom (thiol ether bond) to the protein. The so-called hedgehog proteins, which are important in cellular development, are modified covalently by both cholesterol and N-palmitoyl moieties.

It is now clear that such modifications are important in determining the activities of proteins and in targeting them to specific subcellular membrane domains, including the rafts in plasma membranes. Thus, both myristoylated and palmitoylated proteins are targeted to rafts (as are the GPI-anchored proteins), but prenylated lipids are not. It is noteworthy that many signalling proteins are modified by lipids with implications for the relevant events at cell surfaces. Lipoproteins are complex aggregates of lipids and proteins (not bound covalently) that render the lipids compatible with the aqueous environment of body fluids and enable their transport throughout the body of all vertebrates. Within the circulation, these aggregates are in a state of constant flux, changing in composition and physical structure as the peripheral tissues take up the various components before the remnants return to the liver. The most abundant lipid constituents are triacylglycerols, free cholesterol, cholesterol esters and phospholipids (phosphatidylcholine and sphingomyelin especially), though fat-soluble vitamins and anti-oxidants are also transported in this way. Free (unesterified) fatty acids and lysophosphatidylcholine are bound to the protein albumin by hydrophobic forces in plasma. Ideally, the lipoprotein aggregates should be described in terms of the different protein components or apoproteins (or apolipoproteins), as these determine the overall structures and metabolism, and the interactions with receptor molecules in liver and peripheral tissues. However, the practical methods that have been used to segregate different lipoprotein classes have determined the nomenclature. Thus, the main groups are classified as chylomicrons (CM), very-low-density lipoproteins

(VLDL), low-density lipoproteins (LDL) and high-density lipoproteins (HDL), based on the relative densities of the aggregates on ultracentrifugation.

Other Lipids
Of course many other lipids occur in nature with important biological functions. For example, lipopolysaccharides, fat-soluble vitamins, rhamnolipids and so forth are not discussed in this document. Similarly, there are many lipids that are unique to specific organisms from bacteria to marine invertebrates that are not discussed above. The reader will find information on the chemistry, biochemistry and functions of most of these in other pages on this site.

1. Introduction
Comprehensive discussion of the enormous literature on compositions of animal, plant and microbial tissues would be a daunting task, and it is only possible here to summarize briefly some of the more significant features of lipid composition, with some highly selective (and simplified) examples of relevant analyses. Further data are available for specific lipids in most of the webpages in this part of the website. One problem in comparing data from different sources is the method of presentation, which is often dependent on the nature of the analytical methods used. For example, it is easier technically to analyse the phospholipids and glycolipids following isolation as distinct groups, separately from the simple lipids. Often there is no attempt subsequently to integrate the data. To add to the confusion, results of analyses of simple lipids are often reported in terms of weight percent of each lipid class, since the data are acquired in this form, while the results for phospholipids are most frequently recorded as molar percent, especially when phosphorus analysis is used as the means of quantification. Many of the data listed below are from publications that may appear old but are nonetheless reliable. The results from modern lipidomics analyses are often difficult to summarize in a simple tabular form.

2. Lipid Class Compositions

In animal tissues, the structural lipid components, such as the phospholipids, tend to be rather constant in composition under the normal physiological conditions so meaningful comparisons between different organs can often be made. On the other hand, the proportions of the simple lipids, especially the triacylglycerols, can vary greatly according to the dietary or physiological state of the animal, and this information is not always recorded in papers. Where possible, data are presented for the liver, because it contains all of the important lipid classes, and because of the central importance of this

organ in lipid metabolism. Marine lipids are discussed only briefly here; Ackman has edited a two-volume work on the subject [1]. As examples, some data on the lipid composition of selected tissues from the rat (an important animal for biological experimentation) are shown in Table 1. Those lipids listed are by far the most abundant in these tissues, and they are those most often seen in other organs, although other lipids can assume importance in some circumstances. Lipids present at relatively low concentrations only, such as most of the lysophospholipids, ceramides, sphingosine, sphingosine-1-phosphate were not determined in these analyses, but they are of course of great metabolic importance. Nor were the diacyl, alkylacyl and alkenylacyl forms of the phospholipids distinguished.
Table 1. The composition of the lipid classes (weight % of the total) in rat heart, liver, erythrocytes and plasmaa.

Lipid Class

Tissue heart liver erythrocytes plasma 16 49 6

trace

trace 2 cholesterol esters triacylglycerols 4 7 cholesterol 4 5 30 trace diacylglycerols 1 trace free fatty acids b cardiolipin 12 5 phosphatidylethanolamine 33 20 21 phosphatidylinositol 4 4 3 phosphatidylserine 3 phosphatidylcholine 39 55 32 sphingomyelin 2 2 8 lysophosphatidylcholine 1 a Christie, W.W., J. Lipid Res., 26, 507-512 (1985). b also contains cerebrosides and phosphatidylglycerol

2 24 2 1

Cholesterol esters, triacylglycerols and free (unesterified) cholesterol tend to be the most abundant simple lipids; when substantial amounts of diacylglycerols and free fatty acids, especially the latter, are encountered in a sample, it is usually indicative of artefactual hydrolysis during storage or extraction of the tissues. The choline-containing constituents, i.e. phosphatidylcholine and sphingomyelin, but predominantly the former, tend to be the most abundant phospholipids (50 to 60% of the total), followed by phosphatidylethanolamine, and then by phosphatidylinositol and phosphatidylserine. The results quoted for rat liver lipids are therefore typical of this and many other organs. Cardiolipin is a major component of mitochondrial lipids in particular and so is found in appreciable amounts in heart muscle; phosphatidylglycerol only appears to assume importance in lung surfactant. Again, when appreciable amounts of phosphatidic acid or lysophospholipids are encountered in samples, artefactual hydrolysis may have occurred during processing or analysis.

In erythrocytes, all the lipids appear to be constituents of the membranes, and the results of the analysis listed here indicate that only cholesterol and the phospholipids are present. These data would be typical of many other species, and any departure from this general pattern is symptomatic of an underlying metabolic difference; for example in ruminant erythrocytes, there is virtually no phosphatidylcholine, which is replaced entirely by sphingomyelin. The fatty acid components of erythrocytes are often considered a better marker of long-term nutritional status than are the plasma lipids. The compositions of the plasma lipids are of particular importance as they supply fatty acids to all tissues, and are assumed to be in some form of compositional equilibrium with them. Of course, biopsy samples of plasma are easy to obtain. Those lipids reported as plasma lipid constituents here are typical; phosphatidylcholine is present as a higher proportion of the total phospholipids than in any other tissue, while cholesterol esters tend to more abundant than in most organs (other than in steroidogenic tissues such as the adrenals). The proportion of triacylglycerols in the plasma in this instance was higher than normal, because an old obese rat was used for the analysis. Lipids are transported in plasma in the form of complexes with proteins, i.e. lipoproteins, which render them compatible with their aqueous environment. It should be recognized that an analysis of the total lipids in plasma can give only part of the picture, and it may be necessary to determine the compositions of each of the lipoprotein fractions before definitive metabolic conclusions can be drawn. All membranes (indeed each side of the bilayer) in a tissue can have distinctive compositions that are in some way related to their function. The results of some analyses of the phospholipids of the membranes of rat liver are recorded in Table 2. Naturally, the same phospholipids are present as is described above for the intact organ, but the relative proportions vary markedly. For example, cardiolipin and sphingomyelin are much more abundant in mitochondria and plasma membrane, respectively, than in any of the other membranes.
Table 2. The phospholipid composition of whole tissue and membrane preparations from rat liver (mol % lipid phosphorus).

Lipid class

Membrane Whole Plasma Nuclei Mitochondria Microsomes tissue membrane [a] [b] [c] [c] [c] 26 15 34 2 22 20

cardiolipin 5 phosphatidylethanolamine 25

phosphatidylinositol 7 4 7 8 7 phosphatidylserine 3 6 1 4 4 phosphatidylcholine 51 57 41 59 43 sphingomyelin 4 6 2 4 23 lysophosphatidylcholine 1 1 2 2 References: a, Wuthier, R.E. J. Lipid Res., 7, 544-550 (1966). b, Gurr, M.I. et al. Biochim. Biophys. Acta, 106, 357-370 (1965). c, Colbeau, A. et al. Biochim. Biophys. Acta, 249, 462-492 (1971). The data listed in Table 2 are incomplete, however, as cholesterol is an important constituent of membranes, modifying their fluidity, and the proportion relative to the phospholipids should be determined. In plasma membrane preparations from rat liver, the molar ratio of cholesterol to phospholipids is 0.76, while that in microsomes and mitochondria is 0.1 [2]. In addition, only a part of each phospholipid is the diacyl form and in most tissues, the phosphatidylethanolamine especially tends to contain an appreciable proportion of plasmalogens. On the other hand, the plasmalogen form of phosphatidylcholine appears only to occur in high proportions in heart muscle and reproductive tissues. With a few significant exceptions, the lipid compositions of equivalent membranes from different tissues and even from different species are in general rather similar, probably because these membranes have common functional requirements (though the fatty acid components may differ).

3. Fatty Acid Compositions of Glycerolipids

The fatty acid composition of each lipid in a tissue is frequently distinctive and can vary markedly between species. It is obviously greatly, but not entirely, dependent on the nature of the diet of the animal concerned (although in many analytical studies, such details are not given). Kuksis [3] has reviewed the fatty acid compositions of animal glycerolipids. Data for the fatty acid compositions of the depot fats, which are highly responsive to dietary influence, in several animal species are listed in Table 3.
Table 3. The fatty acid compositions (mol % of the total) of the adipose tissue lipids of various animal species.

Fatty acid

Species Rat [a] Pig [b] Sheep [c] 23 5 6 35 19 2 29 3 18 41 8 21 2 35 31 2 -

Horse [a] 26 8 5 31 9 18

Herring [d] 21 11 23 1 10

Seal [d] 10 16 26 2 14

16:0 16:1 18:0 18:1 18:2 18:3 20:1

20:5 9 7 22:1 10 7 22:6 5 8 References: a, Brockerhoff, H. et al. Biochim. Biophys. Acta, 116, 67-72 (1966). b, Christie, W.W. and Moore, J.H. Biochim. Biophys. Acta, 210, 46-56 (1970). c, Christie, W.W. and Moore, J.H. J. Sci. Food Agric., 22, 120-124 (1971). d, Brockerhoff, H. and Hoyle, R.J. Arch. Biochem. Biophys., 102, 452-455 (1963). In all, there are relatively high contents of 16:0 and 18:1 fatty acids. Species differences are most apparent in the relative concentrations of the saturated and polyunsaturated components. In the rat, pig and horse, for example, there are appreciable amounts of linoleic acid; this is a minor component in the sheep, because it is subjected to microbial biohydrogenation in the rumen, and in the herring and seal, because it is present at low levels only in their food chain. Although the sheep and horse may consume a very similar diet, there are high levels of linolenic acid, derived from the herbage, in the lipids of the horse and not in those of the sheep, again because of biohydrogenation in the rumen of the latter. Fish, such as the herring, tend to contain relatively high amounts of C20 and C22 fatty acids, derived from the microflora and microfauna, which they consume. In turn, the composition of the depot fat of the seal reflects its diet of fish. Each lipid class in a tissue has a distinctive fatty acid composition also, probably related to its function but in a way that is still only partly understood. Some data for the main glycerolipids and the cholesterol esters of rat liver are listed in Table 4.
Table 4. The fatty acid compositions (mol % of the total) of the main glycerolipids and the cholesterol esters of rat liver.

Fatty acid

TG [a]

PC [a]

PE [a]

Lipid class PI [b] PS [c]

Cardx [d]

CE [e]

16:0 27 14 18 7 4 7 24 16:1 4 1 8 3 18:0 7 34 37 40 47 4 20 18:1 27 10 8 3 3 20 15 18:2 12 12 5 3 2 59 16 20:4(n-6) 1 20 23 40 25 2 14 22:6(n-3) 4 7 2 16 Abbreviations: TG, triacylglycerols; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PS, phosphatidylserine; Card, cardiolipin; CE, cholesterol esters. x of the mitochondrial fraction References: a, Wood, R. and Harlow, R.D. Arch. Biochem. Biophys., 131, 495-501 (1969). b, Holub, B.J. and Kuksis, A. J. Lipid Res., 12, 699-705 (1971). c, Holub, B.J. and Kuksis, A. Adv. Lipid Res., 16, 1-125 (1978). d, Colbeau, A. et al. Biochim. Biophys. Acta, 249, 462492 (1971). e, Connellan, J.M. and Masters, C.J. Biochem. J., 94, 81-84 (1965).

In essence, only C16 and C18 fatty acids are found in significant amounts in the triacylglycerols, although most of the glycerophospholipids contain substantial proportions of the longer-chain polyunsaturated components. For example, the phosphatidylcholine contains 50% of saturated fatty acids, while arachidonic acid constitutes 20% of the total. The phosphatidylethanolamine has a similar proportion of saturated fatty acids, but somewhat less linoleic acid and correspondingly more of the C20 and C22 polyunsaturated fatty acids. Characteristically, high proportions of stearic and arachidonic acids are present in the phosphatidylinositol, while the composition of the phosphatidylserine is similar except that the 22:6(n-3) fatty acid substitutes for part of the arachidonic acid. The cardiolipin differs markedly from all the other glycerophospholipids in that the single fatty acid, linoleic acid, comprises nearly 60% of the total. The composition of the cholesterol esters tends to resemble that of the phosphatidylcholine, but there is somewhat less stearic and more palmitic acid in the former. Similar general compositional trends are seen, although the absolute values may differ, in comparing the same lipids in other tissues and, with dietary influences superimposed, in comparing the corresponding lipids of other species.

4. Fatty Acid Distributions within Glycerolipid Molecules

In addition to each lipid class in a tissue having a distinctive fatty acid composition, each position of the glycerol moiety tends to have a unique fatty acid composition that is determined during the biosynthesis of a lipid mainly by the specificities of various acyltransferases. Again, the positional distributions of fatty acids in particular lipid classes can vary markedly between tissues and species and can have some metabolic importance. Data for triacylglycerols [4] and for glycerophospholipids [5] have been reviewed. Some results for the principal glycerolipids of rat liver, taken from a single systematic analytical study [6], are listed in Table 5.
Table 5. Positional distributions of fatty acids in each position (sn-1, 2 or 3) of the principal glycerolipids of rat liver (results are expressed as mol % of the total in each position) [6].

Fatty acid

Lipid class and position phosphatidylphosphatidyltriacyl-sn-glycerol choline ethanolamine sn-1 sn-2 sn-3 sn-1 sn-2 sn-1 sn-2 57 4 9 22 4 13 3 4 58 19 1 12 4 8 61 11 1 23 1 65 7 1
trace

16:0 16:1 18:0 18:1 18:2 20:4(n-6) 22:6(n 3)

6 1 4 13 23 39 7

25
trace

11
trace

65 8 -

8 8 10 46 13

In the triacylglycerols, the highest proportion of the 16:0 is in position sn-1, but the other saturated fatty acid, 18:0, is distributed between positions sn-1 and sn-3 mainly. The 16:1 is found in equal abundance in all three positions, but the C18 unsaturated fatty acids are distributed mainly between positions sn-2 and sn-3. In the phosphatidylcholine, much of the saturated fatty acids are in position sn-1, although a significant relative proportion of the 16:0 is also found in position sn-2. Some of the 18:1 is in position sn-1, but virtually all of the remaining unsaturated fatty acids are in position sn-2. Similar relative distributions are seen in the phosphatidylethanolamine, except that a slightly greater proportion of the saturated fatty acids are present in position sn-2. 1,2-Diacyl-snglycerols are a common intermediate in the biosynthesis of each of these lipid classes, but there is little correspondence in the compositions of this part of the molecules, when the phospholipids are compared with the triacylglycerols. In addition, analyses of the molecular species distributions in the three classes of lipids indicated that there was little in common between the structures of the triacylglycerols and those of the phospholipids [6]. This suggests that there is great selectivity for particular molecular species of the intermediate diacylglycerols, utilized for the synthesis of each lipid class, or that some hydrolysis and re-acylation of lipids occurs to give the final compositions (the "Lands' cycle").

Much of the published work on the structures of animal triacylglycerols has been concerned with those of adipose tissue, which tends to contain most of the body stores of fat in the form of this single lipid class. The results quoted here for rat liver triacylglycerols are not dissimilar to those obtained for adipose tissue for many animal species. In most instances, position sn-1 contains appreciable amounts of the saturated fatty acids, position sn-2 contains mainly unsaturated and any shorter-chain fatty acids, and position sn-3 consists predominantly of unsaturated and longer-chain fatty acids. The principal exception to this type of distribution was thought to be the triacylglycerols of the pig and related species, in which position sn-2 is occupied largely by palmitic acid (70% or more), but increasing numbers of tissues containing triacylglycerols with a structure of this kind are being revealed as research progresses. For example, milk fats from all the higher mammals studied have more than half of their total content of palmitic acid in position sn-2, and the same is true of the triacylglycerols of lymph, plasma and adrenals in ruminant animals. Another interesting feature of milk fats, especially those of ruminants, is that all the short-chain fatty acids (4:0 and 6:0 specifically) are located exclusively in position sn-3. A number of excellent analytical studies of the molecular species distributions in animal triacylglycerols have been published, but as the results are not easily summarized, readers are referred to a specialist review for detailed information [4].

The positional distributions of fatty acids in each of the two most abundant phospholipids of rat liver, as listed in Table 5, are typical for these lipids in many other tissues and species. In some tissues, the proportions of saturated fatty acids in position sn-2 can be somewhat higher, but polyunsaturated fatty acids never appear to occur in appreciable amounts in position sn-1. Perhaps the best-known example of a glycerophospholipid containing a high proportion of saturated fatty acids is the phosphatidylcholine of lung surfactant, of which up to 60% can be the dipalmitoyl form. The distribution of fatty acids in phosphatidylinositol follows the common pattern, and a high proportion consists of the 1-stearoyl-2-arachidonoyl molecular species.

5. Sphingolipid Compositions
The sphingolipid components of many tissues and species have been analysed, but those of bovine kidney have been studied in detail with respect both to the relative amounts in various regions of the organ and to the nature of the lipid and non-lipid portions of the molecules [7,8]. A few of the results obtained are listed in Table 6 for illustrative purposes.
Table 6. The sphingolipid composition of different bovine kidney regions (results expressed as mg lipid per gram dry tissue) [7].

Lipid class ceramides glucosylceramides galactosylceramides di- and triglycosylceramides sulfatides sphingomyelins

Cortex 1.0 0.6 0.2 0.3 0.1 17.8

Medulla 0.7 0.6 1.3 0.7 0.9 9.8

Whole tissue 0.8 0.4 0.4 14.4

Sphingomyelin is by far the most abundant sphingolipid, but significant amounts of ceramides and mono-, di- and triglycosylceramides, and sulfatides are present also. The relative proportions of each vary appreciably at different sites in the kidney, presumably because of the differing functions of the membranes in each region. The fatty acid composition of the sphingomyelin fraction of rat liver is listed in Table 7 [9]. In comparison to the glycerophospholipids, a very different range of fatty acids is seen. For example, virtually no long-chain polyunsaturated fatty acids are found in sphingomyelin (and other sphingolipids, although they are sometimes incorrectly identified as constituents), but C20 to C24 saturated and monoenoic fatty acids, with odd or even chain-lengths, are present in addition to the normal C16 and C18 components. A similar range of fatty acid constituents is seen in most other sphingolipids, and the bimodal chain-length distribution often causes individual sphingolipids separated by

chromatographic means to appear as two adjacent bands. In addition, sphingoglycolipids frequently contain a similar range of fatty acids to that listed, but with a hydroxyl substituent in position 2.
Table 7. The fatty acid composition (weight % of the total) in the sphingomyelin of rat liver, and the long-chain base composition (weight % of the total) of ceramide from human liver.

Rat liver sphingomyelin Human liver ceramide [7] [9] fatty acid % long-chain base

trace 16:0 22 d16:0 trace 18:0 10 d17:0 18:1 4 d16:1 5 18:2 2 d18:0 2 20:0 2 d17:1 3 a 22:0 14 d18:1(i) 2 23:0 9 d18:1 77 24:0 24 d18:2 8 24:1 13 d19:1(ai) 3 a abbreviations: i, iso-methyl branch; ai, anteiso-methyl branch

The long-chain base composition of ceramide, which is probably similar to that of sphingomyelin, from human liver is also listed in Table 7. Sphingosine accounts for 77% of the total, but a number of other bases are present in small amounts. Relatively simple base compositions of this kind are found in the other sphingolipids of tissues from most simple-stomached animals. In ruminants, on the other hand, the long-chain base compositions can be much more complex, and may include trihydroxy bases.

6. Lipidomic Analyses
As cautioned earlier, it is not easy to present the results of modern lipidomics analyses in a concise tabular fashion. However, it would be remiss not to point readers to a paper from the LipidMaps consortium, which I suspect is destined to become a classic, i.e. an analysis of the lipids of macrophages [10]. It shows what can be achieved when a large dedicated team tackles a problem with modern mass spectrometric methodology. Information on the techniques of lipidomics is available in our webpages here..

Lipid Compositions of Plants and Microorganisms

1. Introduction

Comprehensive discussion of the enormous literature on compositions of animal, plant and microbial tissues would be a daunting task, and it is only possible here to summarize briefly some of the more significant features of lipid composition, with some highly selective (and simplified) examples of relevant analyses. Further data are available for specific lipids in most of the web pages in this part of the website. One problem in comparing data from different sources is the method of presentation, which is often dependent on the nature of the analytical methods used. For example, it is easier technically to analyse the phospholipids and glycolipids following isolation as distinct groups, separately from the simple lipids. Often there is no attempt subsequently to integrate the data. To add to the confusion, results of analyses of simple lipids are often reported in terms of weight percent of each lipid class, since the data are acquired in this form, while the results for phospholipids are most frequently recorded as molar percent, especially when phosphorus analysis is used as the means of quantification. Many of the data listed below are from publications that may appear old, but the results from modern lipidomics analyses are often difficult to summarize in a simple tabular form.

2. Lipid Class Compositions of Plant Tissues

The compositions of the lipids of plant tissues have been reviewed [1]. Some results are listed in Table 1. Those plant tissues that serve as major food materials have received most study obviously. Triacylglycerols tend to be the most abundant class of storage lipid in tissues that are rich in lipids, such as the commercially important oil seeds. On the other hand, there are exceptions and jojoba oil, for example, consists mainly of wax esters. Many storage tissues in plants have starch as the main constituent rather than lipid, and in potato tubers and apples the complex glycolipids and phospholipids are the only lipids present at low levels. In addition to those lipids listed, sterols, sterol esters, acylated sterol glycosides, phytoglycolipid, ceramide, glucosylceramide, phosphatidic acid, Nacylphosphatidylethanolamine, and phosphatidylserine, amongst others, may be found. It should be recognized that seeds and tubers do not have a homogeneous lipid distribution, the endosperm, germ, bran and other organelles each having a distinctive composition.
Table 1. The lipid class compositions (weight % of the total lipids) of various plant tissues.

Lipid class monogalactosyldiacylglycerol digalactosyldiacylglycerol sulfoquinovosyldiacylglycerol triacylglycerol phosphatidylcholine phosphatidylethanolamine phosphatidylinositol

Potato Apple Soybean Clover Rye tuber fruit seed leaves grass 6 16 1 15 26 13 6 1 5 1 5 23 11 6
trace trace trace

Spinach cloroplasts 36 20 5 7 3 2

46 28 4 7 5 1

39 29 4 10 5 2

88 4 2 2

phosphatidylglycerol others Reference

1 15 [2]

1 42 [3]

trace

5 [4]

6 3 [5]

7 4 [1]

[6]

The glycosyldiacylglycerols, i.e. mono- and digalactosyldiacylglycerols and sulfoquinovosyldiacylglycerol, are the most abundant lipid classes in leaf (photosynthetic) tissues. Glycerophospholipids, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylglycerol, are also present and other complex lipids are occasionally reported. Phosphatidylglycerol appears to be especially characteristic of photosynthetic tissue, and it can be the main glycerophospholipid in certain green algae. Triacylglycerols are virtually absent from leaves. As in animal tissues, each of the membranes or organelles in the leaf has a characteristic lipid composition. Spinach chloroplasts have received a great deal of attention, because they can be prepared relatively easily for biochemical experiments, and like the intact leaf they contain appreciable amounts of the glycolipids and a smaller proportion of glycerophospholipids. In contrast, as in animal tissues, the plasma membrane has a high content of phosphatidylcholine, while the mitochondria contain cardiolipin. A further region of plants with a distinctive composition is the epidermis or cuticle. The lipids here tend to be rich in waxes, and can include cutin and suberin, which are complex polyesters of hydroxy fatty acids. Leaf surface lipids are largely non-polar waxes. It has been argued that these may be the most abundant lipids on Earth, considering that leaves cover much of the land surface.

2. Fatty Acid Compositions of Plant Tissues

The fatty acid compositions of the seed oils of importance to commerce have been reviewed [7], and data for a few typical ones are listed in Table 2. Maize (corn), sunflower and safflower oils are of nutritional value since they contain appreciable amounts of the essential fatty acid - linoleic acid. Excessive amounts of linolenic acid, as in soybean oil, can lower the commercial value of an oil because it is then more susceptible to rancidity problems caused by autoxidation; it is therefore a common industrial practice to subject the oil to hydrogenation. In contrast, there are no such problems with olive oil, an important lipid constituent of the Mediterranean diet, with its high content of oleic acid. Palm oil contains a higher proportion of saturated fatty acids than most seed oils. Similarly, cocoa butter consists largely of molecular species with saturated fatty acids in positions sn-1 and 3 and oleic in position sn-2. Rapeseed is one of the few oil crops capable of being grown in northerly climates. In its native form, it tends to have a high content of erucic acid (22:1(n-9)), which may have some properties that may be harmful to the consumer, although this is still a matter for

controversy. However, new cultivars with negligible levels of this component (Canola) are now widely grown. Cottonseed oil resembles maize oil in its composition, but also contains small amounts of the cyclopropene fatty acid, sterculic or 9,10methyleneoctadecenoic acid, which has well-established toxic properties and must be removed during refining. Palm kernel and coconut oils are noteworthy for a high content of saturated fatty acids of medium chain-length.
Table 2. The fatty acid compositions (weight % of the total) of some seed oils [7].

Fatty acid Soybean Maize

Seed oil Safflower Rapeseeda 3 1 11 13 9 55

Olive 12 2 72 8 1

Palm 42 4 38 9

16:0 11 11 6 trace 18:0 4 3 18:1 23 25 12 18:2 51 57 73 18:3 7 1 1 C20-C22 a newer cultivars can contain much less erucic acid

The picture will become more complex as new genetically modified seed oils are introduced. In addition, there are many seed oils which may have limited or negligible commercial value at present, but contain fatty acids with unusual substituent groups and are of great interest to biochemists. Each of the lipids in a plant tissue can have a characteristic fatty acid composition and for illustrative purposes, some results for spinach leaf lipids are listed in Table 3.
Table 3. The fatty acid compositions (weight % of the total) of the individual lipids of spinach leaves [8].

Fatty acid MGDG

a

DGDG

Lipid class SQDG PG

PC

PI

PE 46

trace 16:0 6 27 22 20 41 trace trace 16:1(3t) 35 trace 16:3 30 3 trace 18:0 1 1 18:1 1 4 6 2 11 6 18:2 1 3 39 5 30 35 18:3 67 84 28 36 40 27 a MGDG, monogalactosyldiacylglycerol; DGDG, digalactosyldiacylglycerol; SQDG, sulfoquinovosyldiacylglycerol; PG, phosphatidylglycerol; PC, phosphatidylcholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine.

1 2 7 43

Glycosyldiacylglycerols tend to consist mainly of the unsaturated fatty acids, linoleic acid and especially linolenic acid; a hexadecatrienoic (16:3(n-3)) acid may be present also with certain species. On the other hand, the glycerophospholipids contain higher proportions of saturated fatty acids, generally palmitic acid, in addition to the unsaturated components. Phosphatidylglycerol is unique in that it contains a substantial amount of an unusual fatty acid, i.e. trans-3-hexadecenoic acid. The fatty acid compositions of plant tissues can vary with climatic and other cultivation conditions, and with the stage of development of the tissue, and major species differences occur. However, the results listed in Table 3 are typical.

3. Fatty Acid Distributions within Glycerolipids in Plant Tissues

Once more, the triacylglycerols of the commercial seed oils are those to have been subjected most frequently to detailed structural analyses. In general, there tends to be little difference between the compositions of positions sn-1 and sn-3 of the glycerol moiety, but the saturated fatty acids are concentrated in the primary positions and the unsaturated are in greatest abundance in position sn-2. In some instances, there appears to be a higher proportion of longer-chain fatty acids (C20 to C22) in position sn-3 than in position sn-1, and sometimes the more unusual fatty acids are concentrated in position sn3. The positional distributions of fatty acids in many of the glycerophospholipids of plants seem to resemble those of animal tissues in that the saturated fatty acids are concentrated in position sn-1 and the unsaturated in position sn-2. However, phosphatidylglycerol from spinach leaves and the model plant Arabidopsis thaliana, is unusual in that the major molecular species contains linolenic acid in position sn-1 and trans-3hexadecenoic acid in position sn-2 [9,10]. Data for both glycoglycerolipids and glycerophospholipids are listed in Table 4.
Table 4. Composition (mol %) of fatty acids in positions sn-1 and sn-2 of mono- and digalactosyldiacylglycerols, phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol from leaves of Arabidopsis thaliana [9].

Lipid class MGDG

Position sn-1 sn-2 sn-1 sn-2 sn-1 sn-2

16:0 2
trace

16:3 1 70 2 3

Fatty acids 18:0 18:1

trace trace trace trace trace trace

18:2 4 1 3 4 23 47

18:3 93 28 76 83 26 47

DGDG

15 9 42 1

2
trace

PC

4
trace

5 5

PE

sn-1 sn-2 sn-1 sn-2

58
trace

4
trace

5 2 9 1

15 60 13 8

18 38 55 8

PG

22 23

3t -16:1 41

trace trace

In those plants containing 16:3(n-3), the monogalactosyldiacylglycerols consist mainly of the 16:3-18:3 combination (with all the 16:3 in position sn-2), while the digalactosyldiacylglycerols have the more common 18:3-18:3 species. The distinctive compositions of the phosphatidylglycerol and monogalactosyldiacylglycerol are believed to result from the existence of a primitive prokaryotic biosynthetic pathway in addition to the more usual eukaryotic pathway in the chloroplasts [11]. This is discussed further in our web page on glycosyldiacylglycerols.

4. Lipids of Microorganisms

Lipid Class Compositions

It is not easy to generalize about the lipids of microorganisms, as each family tends to have a distinct and characteristic lipid composition, and there are many lipid classes that are unique to particular groups. In addition, the fatty acid components are often very different from those of animal or plant tissues, and there are some that have as yet been found in certain rare species of microbes only. For this reason, I have not attempted to tabulate data here. Microbial lipids have been reviewed elsewhere in great detail [12]. The nature and composition of microbial lipids have proved to be of great taxonomic value, and their study has assisted towards an understanding of the molecular basis of evolution. Simple glycerolipids such as triacylglycerols are often the most abundant lipids in fungi, but bacteria do not normally accumulate storage lipids of this type, although they are known to be present in a few species; diacylglycerols may be formed transiently as intermediates in the biosynthesis of glycerophospholipids. Sterols are found in yeasts, fungi and algae, but in essence are absent from bacterial membranes.

Most of the common glycerophospholipids of plants and animals that are described above are present in microorganisms. For example, phosphatidylethanolamine is often the most abundant lipid class in many bacterial species (including both Gramnegative and Gram-positive bacteria), and may be accompanied by phosphatidylserine, phosphatidylglycerol, and phosphatidylinositol. The N-mono- and N,N-di-methyl derivatives of phosphatidylethanolamine are found in some bacterial genera. On the other hand, phosphatidylcholine is not a ubiquitous constituent of microbial lipids, rarely being found in bacteria, although it is often the major lipid class in eukaryotes. In bacteria, phosphatidylglycerol especially is widely distributed and it is found in most genera; uniquely it is the only glycerophospholipid in the membranes of certain cyanobacteria. Phosphatidylglycerol is usually accompanied by cardiolipin, which is often a major lipid component of bacteria, although it is only found as a constituent of mitochondrial membranes in eukaryotes. In addition, derivatives of phosphatidylglycerol (lipoamino acids) in which the glycerol moiety is esterified to an amino acid, such as lysine, ornithine, or alanine, are common in Gram-positive bacteria. The plant glycosyldiacylglycerols are also found as major constituents of the membranes of algae and cyanobacteria, although glucose may replace a galactose unit in some species. Diglycosyldiacylglycerols, in which the carbohydrate moiety may be two glucose, two mannose or less often two galactose units, are found as minor components of the membranes of Gram-positive and occasionally of Gram-negative bacteria. Similarly, the plant sulfolipid, sulfoquinovosyldiacylglycerol, is present in small amounts in certain photosynthetic bacteria. Other bacterial sulfolipids tend to be sulfate esters of glycolipids. Complex phosphorylceramides containing inositol are major components of yeast lipids, but sphingolipids are rarely found in bacteria. On the other hand, there always appear to be exceptions to any rule of this kind, and ceramide phosphorylethanolamine and ceramide phosphorylglycerol have been detected in some anaerobic bacteria. Free ceramides have been found in some species of Bacteriodes. Aminolipids containing alkylamines, but lacking the hydroxyl groups of the sphingoid bases, occur in some bacteria, while others contain sulfonolipids (capnoids), which appear to be related structurally to the long-chain bases. For example, capnine is 2-amino-3-hydroxy-15methylhexadecane-1-sulfonic acid and is found in the free form or as the N-acyl derivative in some species of gliding bacteria. Many bacterial species contain certain glycerophospholipids and glycolipids (including glycophospholipids), which are apparently found nowhere else in nature. Glycoglycerophospholipids are common constituents of bacterial membranes. For

example, although phosphatidylinositol itself is rarely found, various mannoside derivatives do occur in some species. Glycosylated forms of phosphatidylglycerol are perhaps the most common of all. To add to the complexity, the glyceryl moiety can also be esterified with an amino acid, the nature of which is dependent on the bacterial species, and the stereochemistry of the glyceryl group can vary between species in these lipids. Glucosaminylphosphatidylglycerol has been found in both Gram-negative and Gram-positive bacteria. Certain rumen bacteria contain lipids with complex glycerophosphoryl groups and glycosylglycerol groups, linked by a C32 dicarboxylic acid (diabolic acid), and may span the membrane bilayer. The lipoteichoic acids of the cell walls of Gram-positive bacteria are polymers of glycerol-1-phosphate and other complex organic groups linked to a diglycosyldiacylglycerol.

Relatively simple glycolipids, such as di- and triacylglucoses, and an analogous rhamnolipid are present in some bacteria. Much more complex lipid polysaccharides, for example Lipid A, occur in the cell envelope of certain microbial species with phosphatidylinositol as the anchor moiety in the membrane. Similarly, the cell walls of the Mycobacteria contain a wide range of lipid polymers, including trehalose derivatives, phosphate esters, peptidolipids and phenolic lipids, and these may be esterified to a bewildering array of unusual fatty acids (see below). A further group of distinctive bacterial lipids are the acylornithines. In these, the amine group of ornithine is attached via an amide bond to a fatty acid with a 2- or 3-hydroxyl group, which can in turn be esterified to a further fatty acid. The carboxyl group of the ornithine can also be linked to an aliphatic alcohol. The precise nature of the aliphatic groups and of the linkages can be characteristic of particular species. The glycerophospholipids of bacteria exist not only in the diacyl forms but also as ether lipids, most often the plasmalogen forms. In addition, many different complex di- and tetra-ether phospholipids and glycolipids, in which the aliphatic moieties are phytanyl or related structures, are present in archaebacteria, thermoacidophiles and thermohalophiles [13].

Fatty Acid Compositions

Polyunsaturated fatty acids of the kind found in plant lipids occur also in algae (green and brown), fungi and cyanobacteria, but are not often present in other bacteria (some marine species are exceptions). In general, bacterial lipids tend to contain appreciable amounts of C14 to C18 straight-chain saturated and monoenoic fatty acids. The common C18 monoenoic acid is not oleic acid, however, but cis-vaccenic acid (18:1(n-7)). In addition,

bacterial lipids can contain odd-chain, branched-chain (mainly iso- and anteiso-methyl, but 10-methyloctadecanoic or tuberculostearic acid is characteristic of some species), cyclopropane and 3-hydroxy fatty acids, which are only rarely synthesised by other organisms. The presence of a methyl branch or of a cyclopropane ring in the fatty acids in a membrane increases its fluidity in an analogous manner to that of double bonds in polyunsaturated fatty acids in the membranes of higher organisms. In comparing the detailed fatty acid compositions of bacteria, it is important to recognize that they can vary greatly with culture conditions and with stage of growth. The Mycobacteria and certain related species contain a highly distinctive range of verylong-chain -branched -hydroxy fatty acids. They occur in the bacterial cell walls in the free form, as wax esters and as components of complex lipids.

5. Lipidomic Analyses
As cautioned earlier, it is not easy to present the results of modern lipidomics analyses in a concise tabular fashion, but the immense contribution the newer mass spectrometric methods are making to our understanding of lipid compositions cannot be ignored. However, Ruth Welti discusses plant lipidomics elsewhere on this site. Information on the techniques of lipidomics in general is available in our web pages here..