Biosynthesis of Membrane Lipids

and Steroids
Lecture 1
Berg, Tymoczko and Stryer
Chp. 26
No Problems

CDP diacylglycerol
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 Phospholipid and TAG Biosynthesis
1. Phosphatides are precursor of phospholipids and
triacylglycerols.
2. Phosphatides are synthesized by the successive addition
of two acyl CoAs to glycerol-3-phosphate. Both reactions are
catalyzed by glycerol phosphate acyltransferase.

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1. Triacylglycerols are formed by hydrolysis of the phosphate
by a phosphatase followed and addition of a third acyl CoA by
diglyceride acyltransferase.
2. Both activities are associated in the triacylglycerol
synthetase complex localized in the endoplasmic reticulum
membrane.

*for phospholipid
biosynthesis, one of the
reactants must be activated.
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Phospholipids are synthesized in two ways.
1. From activated diacylglycerol (DAG)
Phosphotidyl inositol:
Phosphatide + CTP CDP-diacylglycerol + PPi
CDP-diacylglycerol + inositol Phosphatidylinositol + CMP
1) subsequently phosphorylated to form the signaling
molecule phosphatidylinositol 4,5-bisphosphate.
2) while most phospholipids contain a variety of different
fatty acids, phosphatidyl inositol contains stearic acid at C1
and arachidonic acid at C2.
Cardiolipin: CDP-diacylglycerol + phosphatidylglycerol
CMP +

cardiolipins are located in the

inner mitochondrial membrane. 4
2. From activated alcohol
Phosphatidylethanolamine
DAG + CDP-ethanolamine Phosphatidylethanolamine
+ CMP
Phosphatidylcholine (most common phospholipid in
mammals) is synthesized in two ways.
choline, activated as for ethanolamine, reacts with
DAG.
phosphotidylethanolamine methyltransferase in liver
adds three methyl groups to ethanolamine.

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Phosphatidylserine
Base exchange reaction
1. phosphatidylcholine + serine phosphatidylserine + choline
2.phosphatidylethanolamine + serine phosphatidylserine +
ethanolamine

In mammals, phosphatidylserine represents 10% of the total

phospholipid, while phosphatidylcholine represents about 30%.

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 Sphingolipids
Sphingolipids are present in all cells but largest amounts in
the central nervous system. May be precursors of second
messengers.
1. Sphingosine is formed from palmitoyl
CoA and serine.
2. The amino group is acylated by a
long chain acyl CoA.

3. The hydroxyl group is substituted,

a. choline = sphingomyelin
b. glucose or galactose = cerebroside 7
 Gangliosides
Gangliosides contain an oligosaccharide chain with at least one
acidic residue attached to the ceramide.
1. The acidic sugar, called a sialic acid, is N-acetyl
neuraminate or N-glycolylneuraminate.
2. Attached in an ordered, stepwise fashion.

GM1

1. Requires activated sugars, UDP-glucose, UDP-galactose,

UDP-N-acetylglactosamine and CMP-N-acetylneuraminate.
2. There are more than 60 varieties determined by the
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specificity of the glycosyltransferases in the cell.
Diseases associated with lipid abnormalities Tay-Sachs
lysosomes
1. Respiratory distress syndrome: defect in
dipalmitoyl phosphatidylcholine synthesis.
The lack of this lipid in the extracellular fluid
surrounding the lungs tends to cause collapse.
2. Tay-Sachs disease: a genetic defect that
prevents degradation of gangliosides which
comprise 6% of the lipid in gray matter of the brain.
The sugars are normally removed sequentially in
the lysosomes. In this disorder, the lysosomes
become swollen with lipid.

very slow
-N-acetylhexosaminidase
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 Regulation of Lipid Biosynthesis
1. Not well understood.
2. PAP (phosphitidic acid phosphatase or lipin-1) in
conjunction with diacylglycerol kinase is involved in
regulating the relative amounts of phosphatidate and
diacylglycerol.
Lipin 1 Activated by CDP-diacylglycerol,
Inhibited by sphingosine and
dihydrosphingosine phosphotidylinositol and cardiolipin

Regulates growth of ER and

nuclear membrane and is Reacts with activated
involved in actiivation of PKC alcohol
genes involve in
phospholipid synthesis. 10
 Cholesterol
Common component of animal cell membranes that
modulates membrane fluidity and serves as a precursor of
steroid hormones.
1. All of the carbons comes from acetyl CoA.
2. Synthesis occurs in three phases.
Stage 1. isopentenyl pyrophosphate synthesis (cytoplasm).
Stage 2. condensation of 6 isopentenyl pyrophosphates to
form squalene (ER).
Stage 3. cyclization of squalene to form cholesterol (ER).

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Stage 1
1. Acetoacetyl CoA and acetyl CoA condense to form 3-
hydroxy-3-methylglutaryl CoA.
2. It has two fates,
a. In the cytosol it is irreversibly reduced to mevalonate,
a cholesterol precursor, the first committed step.
b. In the mitochondrial, it is converted to ketone bodies.

3. Mevalonate formation is catalyzed

by 3-hydroxy-3-methylglutaryl CoA
reductase (HMG-CoA reductase), an
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integral membrane protein in the ER.
4. Mevalonate is converted in four steps to 3-
isopentenyl pyrophosphate.
5. The first three steps require ATP.
6. The last step is a decarboxylation.
3-isopentenyl pyrophosphate is the activated
isoprene unit used in the synthesis of cholesterol.

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Stage 2
1. isopentenyl pyrophosphate isomerizes to dimethylallyl
pyrophosphate.
2. Isopentenyl pyrophosphate and dimethylallyl pyrophosphate
condense to form geranyl pyrophosphate (C10).

3. The same kind of reaction occurs again yeilding C15

farnesyl pyrophosphate.
C5 C10 C15 C30
4. The C15 compounds undergo reductive tail to tail
condensation catalyzed by squalene synthase to form14
the C30 compound, squalene.
Stage 3
1. Squalene is activated by formation of an epoxide.
2. Squalene epoxide then cyclizes to lanosterol in a
reaction catalyzed by oxidosqualene cyclase.

3. The enzyme binds the substrate

in a hydrophobic cavity in the correct
conformation and protonates the epoxide.
4. The carbocation then spontaneously
rearranges to form lanosterol.
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Cholesterol is formed from
Lanosterol in multiple steps.

1. Removal of 3 methyl
groups.
2. Reduction of a double
bond by NADPH.
3. Migration of the other
double bond.

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 Regulation of Cholesterol Biosynthesis
1. Biosynthesis occurs primarily in liver, also intestine.
2. The rate of biosynthesis depends on the cellular level of
cholesterol. Feedback regulated by contolling the activity and
amount of HMG CoA reductase.
3. Regulatory mechanisms
a. The sterol regulatory element
binding protein (SREBP), a
transcription factor that binds
to the sterol regulatory
element (SRE) upstream
of the HMG CoA reductase
gene and other genes involved
in cholesterol biosynthesis.
The binding of SREBP to SRE
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increases gene transcription.
The SREBP is anchored to the ER membrane in association
with SREBP Cleavage Activating Protein (SCAP). SCAP is
the cholesterol sensor. When cholesterol levels fall, SCAP
escorts SREBP to the Golgi where it undergoes two specific
proteolytic cleavages.

SREBP is released
from the membrane
and migrates to the
nucleus and activates
the expression of the
HMG-CoA reductase
gene.
Cholesterol synthesis
increases.

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Cholesterol Low
SCAP binds to vesicular proteins for transport of SCAP-
SREBP to the Golgi.

Cholesterol High
1. The binding of cholesterol to SCAP induces a
conformational change.
2. Now SCAP binds to another ER protein called Insig which
anchors SCAP-SREB to the ER membrane preventing is
transport to the Golgi no activation.
3. Binding of the cholesterol metabolite, 25-hydroxycholesterol
to Insid also promotes it interaction with SCAP. 19
 When cholesterol levels increase, there is no further
degradation of the membrane bound SREBP and the
active transcription factor in the nucleus is rapidly
degraded.
b. The translation of the reductase RNA is inhibited by
metabolites derived from mevalonate and dietary
cholesterol.
c. The reductase turnover is precisely controlled. The
membrane bound domain senses degradation signals.
Increasing sterols alters the conformation and increases
sensitivity to proteolysis. The protease is similarly
controlled. Reductase is also subject to ubiquination and
degradation by the 26S proteosome.
Control mechanisms alter the enzyme level 200-fold.
d. Phosphorylation by an AMP activated protein kinase turns
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off the enzyme. No cholesterol synthesis when ATP low.
Controlled by Degradation of the Reductase
1. The membrane domain of HMG-CoA Reductase binds
sterols (eg. Lanosterol and 25-hydroxycholesterol).
2. Induces an association with Insig that are bound to
ubiquitinating enzyme.
3. The reductase in ubiquinated.
4. Geranylgeraniol induces extraction from the membrane.
5. Released to the cytoplasm where digested by proteosomes.

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 Transport Particles
Cholesterol and phospholipids are transported in the
blood stream and other body fluids as lipoprotein
particles that consist of a core of hydrophobic lipids and
a shell of more polar lipids and proteins. The proteins,
1. Solubilize the hydrophobic lipids.
2. Contain targeting signals.
Lipoprotein particles are classified by density.

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Chylomicrons
Transport dietary phospholipids, cholesterol and other lipids
from the intestine.
1. Large, 75 - 1200 nm in diameter.
2. Density < 0.95 gm/cm3
3. Apoprotein B-48, 240 kDa, forms a amphipathic shell
around the lipid core. Also apolipoprotein C and E
4. The hydrophobic regions are in contact with the lipid, the
hydrophilic regions are on the external face of the particle.
5. The triacylglycerides are released by the lipoprotein
lipases located in the lining of blood vessels near muscle
and other tissues.
6. The remainder rich in cholesterol, the chylomicron
remnants are taken up by the liver. 23
 Very Low Density Lipoproteins (VLDL)
1. Transport triacylglycerols and cholesterol from the liver.
2. Low density < 1.006 gm/cm3.
3. Contain two proteins, apo B-100 (513 kDa) and apo E (34
kDa). Apo B-100 is encoded by the same gene as apo B-
48, by alternate mRNA processing (editing).
4. The triacylglycerols are hydrolyzed by lipases on the
surface of cells.
5. The remnant, the intermediate density lipoprotein,
density < 1.019 gm/cm3, are rich in cholesterol esters.
a. Half are reprocessed by the liver.
b. Half are converted into low density lipoprotein, LDL,
by removal of more triacylglycerols. 24
 Low Density Lipoprotein (LDL)

diameter = 22 nm
mass = 3 Mda
core = 1500 cholesterol ester molecules
1.019 gm/cm3 < density < 1.063 gm/cm3
1. Major carrier of cholesterol.
2. Major fatty acid of the ester is linoleate.
3. A shell of phospholipid and unesterfied cholesterol
surrounds the core.
4. The shell also contains one molecule of apo B-100 that is
recognized by target cells.
5. LDLs transport cholesterol to peripheral tissues and 25
regulate de novo cholesterol synthesis.
 High Density Lipoprotein (HDL)
1. 1.063 < density < 1.21
2. Picks up cholesterol in the plasma from cell
destruction and membrane turnover.
3. Reverse cholesterol transport
4. An acyltransferase in the HDL esterfies the
cholesterol.
5. The cholesterol esters are transported to the liver by
the HDL.

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