Chapter 10 Lipids ()

The fatty substance, separated from salifiable bases,

was dissolved in boiling alcohol. On cooling, it was
obtained crystallized and very pure, and in this state
it was examined. As it has not been hitherto described
I purpose to call it margarine (), from Greek
word signifying pearl (), because one of its characters
is to have the appearance of mother of pearl, which it
communicates to several of the combinations of which
it forms with the salifiable bases.

-Michel-Eugene Chevreul,

article in Philosophical Magazine, 1814
Lectured by Dr. Qin Yongmei (): June 2, 2007
Storage lipids

Structural lipids in membranes

Lipids as signals, cofactors, and pigments

Separation and analysis of lipids
Fatty acids
Triacylglycerols
glycerolphospholipids
Sphingolipids
galctolipids & sulfolipids
glycerol dialkyl glycerol tetraethers
Biological
membranes
Cholesterol
Overview
Steroid
hormones
arachidonate
derivates





Roles of Lipids
Energy storage (fatty acids, triacylglycerols)
Structural elements (phospholipids, cholesterol)
Hormones ( sex hormones e.g. Estrogen)
Enzyme cofactors (coenzyme A)
Electron carriers (coenzyme Q, plastpquinone)
Light-absorbing pigments (carotenoids)
Hydrophobic anchors (dolichols)
Emulsifying agents (bile salts)
Intracellular messengers (phosphatidyl inositol)

Storage lipids
Fatty acids:

- Structure and nomenclature
- Physical and chemical properties

Triacylglycerols:

- Structure
- Function
- Food
Fatty acids
Structure and nomenclature
Basic formula: CH
3
(CH
2
)
n
COOH
Carboxylic acids with hydrocarbon chains of
4-46 carbons
Free FAs are found in trace quantities in cells

FAs are either:
(i) part of a lipid molecule
(ii) complexed to a carrier protein
(e.g. albumin on blood)

Saturated or unsaturated, usually linear
Naturally occurring fatty acids
There is a common pattern in the location of
double bonds:

Unsaturated FA:
9
,
12
,
15


Polyunsaturated FA:

double bonds are never conjugated and are in
the cis configuration

(-CH=CH-CH
2
-CH=CH-)n
Naming of fatty acids
CH
3
-(CH
2
)
7
-CH=CH
2
-CH
2
-CH
2
-CH
2
-CH
2
-CH
2
-CH
2
-CH
2
-COOH
C18

9 10
18:0, stearic acid : octadecanoic acid
18:1 (
9
), oleic acid : octadecenoic acid

18:2 (
9,12
), linoleic acid : octadecadienoic acid
(-6)

18:3 (
9,12,15
), -linolenic acid : octadecatrienoic acid
(-3)
Cis
9
Fatty acids
#C Common Name
12 Lauric Acid
14 Myristic Acid
16 Palmitic Acid
16:1 Palmitoleic Acid
18 Stearic Acid
18:1 Oleic Acid ()
t18:1 Elaidic Acid ()
18:2 Linoleic Acid
18:3 Linolenic Acid
20 Arachidic Acid
20:4 Arachidonic acid
24:0 Ligoceric Acid
Essential
Fatty acids
Essential Fatty Acids
Omega-3 (-3) and omega-6 (-6) fatty acids are unsaturated
"Essential Fatty Acids" . The two EFAs are linolenic (-3) and
linoleic (-6). The 3" and 6" indicate where the first double bond
occurs in the fatty acid molecule.
Example: DHA (docosahexenoic acid, C22:6 ) and
AA (arachidonic acid, C20:4)
are both crucial to the optimal development of
the brain and eyes.
Health consideration:
In modern diets: -6 / -3 ratio = 15:1;
Expected ratio: 2:1 to 4:1 reduced mortality from
cardiovascular disease, suppressed
inflammation in patients with rheumatoid
arthritis (),and decreased
risk of breast cancer.
Fatty acids
Physical and chemical properties
Weakly acidic (pKa = 4.5 - 5), ionized at physiological pH

Saturated FAs (chain length > 8C) are solid at room
temperature. Melting point (m.p.) depends on chain length,
the degree of unsaturation

Cis double bonds disrupt intermolecular packing and
therefore lower m.p;

Polyunsaturated FAs are readily oxidized by exposure to
air.

Fatty acids form micelles.
amphipathic
()
zig-zag
kink
Van der Waals interactions between the acyl chains:
cis double bonds + Tm
Fatty acids form micelles
The assembly of micelles is
driven by the hydrophobic
effect
Non-polar interior can dissolve
hydrophobic compound
What is soap ?
TAG + 3KOH (NaOH) 3RCOO
-
K+ + glycerol
FA chains can dissolve in oils while charged carboxyl
group dissolves in water;

Forms a mixed micelle which can remove oils;

Soap form precipitates with divalent cations
(reduces efficiency);

Detergent (modified FAs) do not precipitate with
calcium, used as better cleaning agents.
Triacylglycerols
Structure:
TAGs are fatty acid
esters of glycerols,
with each TAG made
up of 1 glycerol and 3
FAs.
Adipocyte
Function:

TAGs are used as a stored fuel;

Advantages:
(1) Greater energy yield

FA 9 kcal/g (37 kJ/g)
CHO/protein 4 kcal/g (16 kJ/g)

(2) Lipid hydrophobicity

TAGs coalesce into dense droplets and
exclude water

Triacylglycerols
Triacylglycerols
Example:

In a 70 kg person, the energy reserves are:
100,000 kcal TAGs
25,000 kcal protein (muscle)
600 kcal glycogen
40 kcal glucose

TAGs represent 11 kg of body weight (55 kg glycogen).
TAG stores can last weeks to months, and glycogen stores
exhausted less than one day.
Fatty acids in food: saturated vs unsaturated
Source Lauric/Myristic Palmitic Stearic Oleic Linoleic
Beef 5 26-32 20-25 37-43 2-3
Milk 25 12 33 3
Coconut 74 10 2 7
Corn 8-12 3-4 19-24 34-62
Olive 9 2 84 4
Palm 39 4 40 8
Soybean 9 6 20 52
Sunflower 6 1 21 66
Fatty acid composition of three food fats
Possible risks related to trans fatty acids
4 Major trans FA found in food: 18:1t
(less amount of 16:1t and 18:2t)

4 Sources of tFA: margarine, cakes, cookies,
chips etc.

4 Link tFA to breast cancer
(http://www.sciencedaily.com)

4 Link tFA to increased serum cholesterol
(http://cardiology.medscape.com
http://nutrition.about.com)
Hydrogenation of Natural Unsaturated Fats
Purpose: a commercial chemical process to add more hydrogen to
natural unsaturated fats to decrease the number of double bonds
and retard or eliminate the potential for rancidity ().
C C
H
H
H
H
C C
H H
+ H
2
high temperatures
with metal catalysts
Side effect: cause a large percentage of the natural cis double bonds
to change to trans double bonds.
cis
Dietary fat and health
Mainstream nutritional
science has demonized dietary
fat, yet 50 years and hundreds
of millions of dollars of
research have failed to prove
that eating a low-fat diet will
help you live longer

(Science 2001, 291:2536-2545)

Artificial fats (fat substitutes )
- mixtures of the esters of fatty acids with polyglycerol.
R-[OCH
2
-CH(OR)-CH
2
O]
n
-R
R: fatty acids; n: about 3.
- almost completely metabolized like fats, so they are not
calorie-free;
- the polymerized glycerol moiety is not digested and is excreted
primarily in the urine.
Olestra:
- an artificial fat created from sucrose (replace of glycerol)
and up to eight fatty acids;
- too large to be metabolized and passes through the body
unchanged, but because it acts as a lipid, it can cause
depletion of fat-soluble vitamins.

Polyglycerol fatty acid esters:
Waxes
Esters of long chain fatty acids (C14-36) with
long chain (C16-30) alcohols

High melting points (60-100C)

Energy storage (Plankton, )

Water repellant (birds and plants)
Triacontanoylpalmitate
major component
of beeswax
Structural lipids in membranes

+ Phospholipids

+ Sphingolipids

+ Plant galactolipids and sulfolipids

+ Archaebacteria have unique
membrane lipids

+ Cholesterol
29

Phospholipids

2 Classes of phospholipids (PL)
(i) glycerolphospholipids glycerol backbone
(ii) sphingomyelin spingosine backbone

Glycerolphospholipids
- essential for membrane structure
- most abundant membrane lipids
- no genetic defects in humans

Sphingolipids
sphingomyelin, neutral glycolipid and gangliosides
L-Glycerol-3-phosphate (D-glycerol 1-phosphate),
the backbone of phospholipids
Glycerophospholipid
Saturated FA

Unsaturated FA
phosphatidic acid
Phosphatidylethanolamine ethanolamine
Phosphatidylcholine choline
Phosphatidylserine serine

Phosphatidylglycerol glycerol


Phosphatidylinositol inositol 4,5-
4,5-bisphosphate bisphosphate

Cardiolipin phosphatidyl-
glycerol

Some phospholipids have ether-linked fatty acids
Other Phospholipids
Plasmalogens
found in central nervous system and in heart muscle
vinyl either linkage at glycerol C-1
choline and etanolamine head groups
resistant to phospholipases

Platelet activating factor
active at very low concentration (0.1 nM)
released from leukocytes called basophils
cause platelet aggregation and vasoconstriction
(via release of serotonin from platelets)
involved in smooth muscle contraction inflammation,
allergic response
acetyl group makes it water soluble
Sphingolipids
sphingosine (4-sphingenine)
36

Sphingophospholipids

sphingosine
Fatty acid
phosphate
Head group
2nd large class of membrane
lipids

abundant in tissues of central
nervous system

involved in recognition events
at the cell surface, such as
cell recognition & cell-cell
communication

Ceramide (shingosine + FA) is
the structural unit common to
all sphingolipids
37
Glycolipids
(e.g. cerebrosides and gangliosides)
Various monosaccharides e.g. glucose, galactose
sphingosine
Fatty acid
Carbohydrate
Amphipathic, pack together with phospholipids in bilayers
Similarities between phosphatidylcholine and sphingomelin
Sphingolipids at Cell Surface are Sites
of Biological Recognition
The carbohydrate
moieties define
the human blood
groups.
Phospholipids and sphingolipids are degraded in lysosomes
Inherited human diseases resulting from
abnormal accumulation of membrane lipids
Sphingolipid
storage diseases
Polar lipids of membrane undergo constant turnover;

Sphingolipids degraded in lysosomes;

Deficiency in particular enzyme leads to accumulation
of a sphingolipid or breakdown intermediate ;
High incidence (1/3600 births, 1/28 carriers
defect gene in recessive form);
Mental retardation and early death (2-3 years);
Lipid accumulation causes lysosomes to swell
leading to cellular/tissue enlargement;
Swollen lysosomes in CNS tissue neuronal death.
Sphingolipid
storage diseases

Chloroplasts contain galctolipids and sulfolipids
Galactolipids
Sulfolipids
one or two galactose residue are connected by a
glycosidic linkage to C-3 of a 1,2-diacylglycerol
(1,2-DAG) ;
thylakoid internal membrane () lipid;
they make up 70-80% of the total membrane lipids
of a vascular plant, most abundant membrane lipid
in the biosphere.
Phosphate is missing !
A sulfonated glucose residue is joined to a 1,2-DAG;
Sulfonate bears a negative charge
(similar role of phosphate group in phospholipids).
Archaebacteria contain unique membrane lipids

4 ether linkages
Glycerol dialkyl glycerol tetraethers (GDGT)
8 five-carbon isoprene groups
R
Glycerol dialkyl glycerol tetraethers
4 The archaebacteria live in ecological niches with extreme
conditions-high temperatures, low pH and high ionic
strength;

4 Ether bonds in GDGT are more stable than ester bonds.

4 twice the length of phospholipid and sphingolipid;

4 with polar heads consisting of glycerol linked to either
phosphate or sugar residues;

4 Central carbon of glycerol is in R configuration in
archaebacteria, but in S configuration in the eubacteria
and eukaryotes.
47
Lipids and structure of the plasma
membrane
glycolipid
Polar head
hydrophobic
tail
Integral
protein
cholesterol
To minimize interactions of FA chains (non-
poplar), phospholipids aggregates form a bilayer
structure.
Cholesterol
Steroids:
(i) cholesterol and sterols of plants and fungi
(ii) steroid hormones
(iii) bile salts

Roles of cholesterol in mammals
(i) structural component of plasma membrane and
modulates membrane fluidity
(ii) precursor of steroid hormones and bile acids

Rarely found in plants, never in bacteria

Steroids and lipid vitamines can be classified as
polyprenyl compounds
Lipids as signals, cofactors, and pigments
Phosphatidylinositols act as
intracellular signals

Eicosanoids carry messages to
nearby cells

Steroid hormones carry messages
between tissues

Vitamins A and D are hormone
precursors
Phosphatidylinositols in cellular regulation
P456
Eicosanoids ()
paracrine hormones;
derived from arachidonic acid
3 classes: prostaglandins, thromboxanes and leukotrienes

Prostaglandins:
PGE (ether-soluble); PGF (phosphate buffer soluble)

Thromboxanes:
produced by platelets (thrombocytes) and act in the
formation of blood clots.

Leukotrienes
They are able to lure white blood cells to the site of injury
and bind them to the vessel wall. But, overproduction of
leukotrienes cause asthmatic attacks
Metabolism of Natural C20 Cis Fatty acids
Produces Powerful Eicosanoids.

Arachidonic acid (-6)
Eicosapentaenoic acid (-3)
Cyclooxygenase Lipoxygenase
prostacyclins prostaglandins
thromboxanes
leukotrienes Hydroxy FAs
Eicosanoids
- In the late 1950s, Dr. Bergstrm purified the first
prostaglandins and determined their structures.

-Bergstrms student, Dr. Samuelsson is responsible for
the chemical development including determination of more
significant components (e.g. leukotriene) within system. He
could tell us how this system is built up and help us to
understand the relationship between its various
components.

- Dr. Vanes contribution was the discovery of an important
component of the system-prostacyclin and revealed the
secret mechanism behind the drug, acetylsalicylic acid
(inhibition of the formation of prostaglandins)


The Nobel Prize in Physiology and Medicine 1982
"for their discoveries concerning prostaglandins and
related biologically active substances"
Sune K. Bergstrm
Bengt I. Samuelsson John R. Vane
Sweden
Sweden
United Kingdom
b.1916 b.1934
b.1927
Cyclooxygenase (COX), also
called prostaglandin H
2
synthase
(bifunctional enzyme):

- cyclooxygenase activity;
- peroxidase activity
cyclooxygenase
activity of COX
cyclooxygenase
activity of COX
peroxidase
activity of COX
Aspirin and Ibuprofen belong to Non-
steroidal anti-inflammatory drugs
,NSAIDs .
Ibuprofen inhibits the same step,
probably by mimicking
the structure of the substrate or an
intermediate in the reaction.

Aspirin inhibits the cyclooxgenase
activity of COX by acetylating an
essential Ser residue on the enzyme.
Steroid hormones carry messages between tissues
-Derivates of sterols;

-Play important roles
in gene expression;

-Sex hormones and
hormones from adrenal
cortex;

-Prednisolone and
prednisone are steroid
drug with potent
antiinflammatory
activities.
Plant: use steroids and eicosanoid-like compounds
in signaling
derived from
fatty acid C18:3 (
9,12,15
)
Plant defense
Growth regulator
63
Vitamins
Fat soluble: vitamins A,D,E,K
Long chain polyunsaturates hydrocarbons
attached to 1 or 2 benzene rings
Vitamin A (retinol)

stored in liver
precursor: carotene
deficiency: night
blindness

Vitamin D
formed from precursor in
skin by UV
regulates Ca deposition
deficiency: rickets
Active sugar precursors
Vitamin D
3
production
and metabolism
UV light
1,25-Dihydroxycholecalciferol
Cholecalciferol (VitD
3
)
7-Dehydrocholesterol
2 steps in skin
1 step in liver
1 step in kidney
25
Active VitD
3
Vit D
3
is formed in skin;

1,25-dihydroxy VitD
3
is
biological active;

1,25-dihydroxy VitD
3
regulate gene expression,
e.g. turning on the synthesis
of an intestinal Ca
2+
binding
protein;

Deficiency in VitD
3
Vitamins E and K and Lipid Quinones are
oxidation-reduction cofactors

Vit E: biological antioxidants;

Vit K: undergoes a cycle of oxidation and reduction;
during the formation of active prothrombin;

Warfarin: synthetic compound that inhibits the formation
of active prothrombin;

Ubiquinone (coenzyme Q) and plastoquitone:
isoprenoids that function lipophilic electron carriers
in the oxidation-reduction reactions that drive ATP
synthesis in mitochondria and chloroplasts, respectively.

The Nobel Prize in Physiology and Medicine 1943
"for his discovery of vitamin K"
Henrik Carl Peter Dam Edward Adelbert Doisy
Denmark USA
1895-1976 1893-1986
"for his discovery of of the chemical
nature of vitamin K"
Separation and Analysis of Lipids
Lipid extraction requires organic solvents

Adsorption chromatography separates
lipid of different polarity

Gas-lipid chromatography resolves
mixture of volatile lipid derivates

Mass spectrometry

Specific hydrolysis
chloroform/methanol/water
water
liver
methanol/water
chloroform
Thin-layer
chromatography
adsorption
chromatography
NaOH/methanol
Fatty acyl methyl ester
Gas-liquid
chromatography
HPLC
Determination of the structure of a fatty acid by
mass spectrometry
fatty acyl picolinyl () ester
m/z
Summary
FAs and TAGs:
nomenclature,structures and names of all common FAs;
know the essential FAs;
physical properties;
structure of TAGs;
advantages of TAGs as a fuel source
structure of waxes

Glycerolipids:
chemical structures of all the main phopholipids, including
plasmalogen and platelet activating factor;
nomenclature: you should know the general names (e.g.
phosphatudylcholine), specific names (e.g. 1-palmitoyl-2
-oleoyl-phosphatidylcholine) and abbreviation (PC)
head group charge
phospholipase
Summary
Sphingolipids:
structures of sphingosine, ceremide and sphingomyelin;
families of sphingolipids;
understand glycolipid storage disease;

Galactolipids and Sulfolipids

Glycerol Dialkyl Glycerol Tetraethers (GDGT)

Cholesterol:
structure

Phosphatidylinositol bisphosphate; eicosanoid;
steroid hormones; vitamins D,A,E and K; ubiquinones;
dolichols.