Synthesis of Triacylglycerol & Phospholipids

Triacylglycerol Phosphatidylserine

H O

H C O C (CH2)n CH3
O

H C O C (CH2)n CH3
O

H C O C (CH2)n CH3

glycerol Fatty acids

 Some important phospholipids
characterized by different alcohol groups

• Phosphatidylcholine
• Phosphatidylethanolamine
• Phosphatidylserine
• Phosphatidylinositol

Major components of biological membranes

Also play a role in cell signalling
 1st steps in TAG & PL synthesis are the same

H2C-OH
Glycerol-3-Phosphate
HO-C-H
H2C-O- P
FA-CoA CoA
Acyl-CoA: glycerol phosphate 1-acyltransferase*
*prefers saturated fatty acids

H2C-O-FA1
Lysophosphatidate
HO-C-H
H2C-O- P
1st steps in TAG & PL synthesis are the same

H2C-O-FA1
Lysophosphatidate
HO-C-H

H2C-O- P
FA-CoA CoA
Acyl-CoA: 1-acyl glycerol phosphate 2-acyltransferase*

*prefers unsaturated fatty acids

H2C-O-FA1
Phosphatidate FA2-O-C-H
H2C-O- P
 2 fates of phosphatidate

Phosphatidate
H2C-O-FA1
CTP
Phosphatidate
phosphohydrolase
FA2-O-C-H
PPi
H2C-O- P
Diacylglycerol CDP-Diacylglycerol
H2C-O-FA1 H2C-O-FA1
FA2-O-C-H FA2-O-C-H
H2C-OH H2C-O- CDP
 Synthesis of Triacylglycerol

Phosphatidate Diacylglycerol Triacylglycerol

H2C-O-FA1 H2C-O-FA1 H2C-O-FA1
FA2-O-C-H FA2-O-C-H FA2-O-C-H
H2C-O- P H2C-OH FA3
H2C-O-
 Synthesis of phospholipids
CTP
Phosphatidate
PPi

Diacylglycerol CDP-Diacylglycerol

Phosphotidylcholine Phosphotidylethanolamine Phosphotidylinostitol

Phosphotidylserine
 Summary
 The synthesis of both triacylglycerol and
phospholipids starts with the conversion of Glycerol-
3-Phosphate to phosphatidate
 Phosphatidate can be converted into either
diacylglycerol or CDP-diacylglycerol
 CDP-diacylglycerol can be converted into
phophatidylinositol
 Diacylglycerol is converted into either triacylglycerol
or phosphatidylcholine or phosphatidylethanolamine
 Phosphatidylserine is made by replacing the choline
or ethanoline group with serine in the preformed
phospholipid
 Lipid Transport

Adipose Tissue
Intestine

Liver Muscle
 Cholesterol

cyclopentanoperhydro-
phenanthrene nucleus

OH
 Lipid Transport

Intestine: lipids derived from diet

transported as lipoproteins
Liver: lipids made or stored in liver
transported as lipoproteins
Adipose tissue releases fatty acids into
the blood stream (non-esterified fatty
acids (NEFA)
 Lipid Transport
1) Non-esterified fatty acids (NEFA) bound to albumin

FA FA
FA
FA

cholesterol
ester
E PL
E cholesterol
E
protein
2) Lipoproteins
E TAG
 Components of Lipoproteins
• Proteins
– Apolipoproteins (apo)
• Phospholipids
– Phosphatidylcholine most abundant
• Triacylglycerol
• Cholesterol
– Free cholesterol at surface
– Cholesterol ester (attached to fatty acid) in core
 Lipoprotein Classes
• Lipoprotein classes differ in
– protein/lipid ratio
– phospholipid/cholesterol/cholesterol ester/
triacylglycerol ratio
– specific proteins present
• This leads to physical differences in
– size
– density
– charge
Major Lipoprotein Classes
Density (g/ml)
Water 1.000
Chylomicrons <1.006
VLDL <1.006
IDL 1.006-1.02
LDL 1.020-1.060
HDL 1.060-1.210
 Composition of major human
lipoproteins
TAG Chol Chol Ester P’Lipid Protein
Chylos 90 2 3 3 2
VLDL 60 4 16 16 6
IDL 20 8 32 32 18
LDL 7 8 42 22 21
HDL2 5 5 20 26 44
HDL3 2 3 15 28 52
 Apolipoproteins
• Diverse group of proteins that associated with
lipoproteins
• Wide ranges of sizes
– apoB100 – MW 550,000
– apoCI – MW 6,600
• While some (apoB48 & apoB100) remain
associated with same lipoprotein particle, most
(apos A,C,E) can transfer between lipoprotein
particles
 Function of apolipoproteins
• Structural
– Hold particle together
– e.g. ApoB48 & ApoB100
• Enzyme activators or inhibitors
– e.g. Apo CII – activates lipoprotein lipase/
ApoCIII - inhibits
lipoprotein lipase
• Receptor recognition
– Recognised by receptors on the surface of cells
– e.g. ApoB100 recognised by LDL receptor
 Major apolipoproteins

Lipoproteins Major Apolipoproteins

chylomicrons B48,E,AI,AII,CI, CII,CIII
VLDL B100,E, AI,AII,CI,CII,CIII
IDL B100,E
LDL B100
HDL AI,AII,CI,CII,CIII
Lipoprotein Synthesis & Secretion

LDL
Endogenous Pathway
IDL

E
N
Chylomicrons

I
ST
VLDL

TE
IN
Remnants
Exogenous Pathway
LIVER
Nascent HDL
HDL3 HDL2
Reverse Cholesterol Transport Pathway
 Lipoprotein metabolism
Exogenous Pathway

via lymph
INTESTINE chylomicron

capillary Energy in
muscle

Lipoprotein chylomicron F.A

Y

Lipase
TAG in
adipose tissue

chylo
remnant LIVER
 The Exogenous Pathway
Transport of dietary lipids
 Dietary lipids are absorbed into the enterocytes
lining the small intestine
 Phospholipid, Cholesterol, Cholesterol Ester &
TAG are incorporated with apolipoprotein (apo)
B48 to form chylomicrons
 Over 90% of the mass of a chylomicron is TAG.
This means that these very large particles have a
very low density
 Chylomicrons are secreted into the lymph and
then into the blood stream
 The Exogenous Pathway
Transport of dietary lipids cont.
 In the capillaries of adipose tissue and muscle the
chylomicrons become anchored to the enzyme
lipoprotein lipase (LPL)
 LPL hydrolyses the TAG and releases fatty acids
which are taken up by the tissue
 When most of the TAG has been broken down the
particle is released back into the circulation as a
chylomicron remnant
 The cholesterol- rich remnant is removed from the
circulation after binding to receptors in the liver
Lipoprotein metabolism
Endogenous Pathway

VLDL LIVER

Energy in
muscle

Lipoprotein

Y
F.A VLDL Lipase

TAG in
adipose tissue Hepatic
IDL
Lipase
50%
50%

LIVER LDL Peripheral

80% 20% Tissues
 The Endogenous Pathway
Transport of lipids from the liver
 In the hepatocytes of the liver phospholipid,
cholesterol, cholesterol ester and triacylglycerol
combine with apo B100 to form Very Low Density
Lipoprotein (VLDL)
 Over 60% of the mass of VLDL is TAG. Thus, these
particles have a low density but are more dense than
chylomicrons
 VLDL are secreted directly into the blood stream
 VLDL interact with LPL in exactly the same way as
chylomicrons.
 The Endogenous Pathway
Transport of lipids from the liver cont.
 When most of the TAG has been hydrolyzed the
particle is released as Intermediate Density
Lipoprotein (IDL)
 IDL is further converted to cholesterol-rich Low
Density Lipoprotein (LDL)
 LDL is removed from the circulation after
interaction with specific cell surface receptors
 80% of LDL is removed by the liver and 20% by all
other tissues
 Lipoprotein metabolism
Exogenous Pathway Endogenous Pathway

via lymph
chylomicron VLDL LIVER
INTESTINE

capillary Energy in
muscle

Lipoprotein

Y
Lipoprotein chylomicron F.A VLDL Lipase
Y

Lipase
TAG in
adipose tissue Hepatic
IDL
Lipase
50%
50%
chylo
remnant LIVER LDL Peripheral
80% 20% Tissues
 Comparison of Exogenous &
Endogenous Pathways
Exogenous Endogenous
Tissue of origin Intestine Liver
Particle Chylomicron VLDL
Site of secretion Lymph/Blood Blood
Form of ApoB apoB48 apoB100
Remnant Chylo Remnant IDL/LDL
Site of uptake Liver Liver/Peripheral
Receptor LRP/LDL LDL
Ligand apoE apoE/apoB100
 Newly-formed HDL is disc-shaped

Discoidal HDL

-LCAT C = cholesterol
| | | |
CC C C

LCAT = Lecithin:Cholesterol-Acyl Transferase

LCAT
Cholesterol + Phospholipid → Cholesterol Ester + Lysophospholipid
(Cholesterol + FA) (Phospholipid – FA)
Conversion of discoidal HDL to spherical HDL
Discoidal HDL

-LCAT
| | | |
-C CC C C
-C

-C
-C

CE
LCAT- CE CE CE -C
CE
-C
-C

-C

HDL3
 HDL3 continues to accumulate
Cholesterol Ester and becomes HDL2

HDL3

Tissue
cholesterol

HDL2

LIVER
 Cholesterol is removed from HDL2 by the liver
through an interaction with cell surface receptors.
When it has lost most of its cholesterol ester it is
released back into the circulation as HDL3

HDL2 HDL3

CE
CE CE
LIVER CE
CE

CE
 Reverse Cholesterol Transport
Transport of cholesterol from tissues to the liver
• Components produced by both the liver and
intestine come together to form Discoidal HDL
• Discoidal HDL rich in protein, phospholipid and
free cholesterol. It’s major protein is apoA1
• Discoidal HDL interacts with cells and picks up
cholesterol which is then esterified by LCAT
• This converts the particles into spherical HDL3
• Spherical HDL3 continues to accumulate
cholesterol and is converted into HDL2
• The excess cholesterol ester is removed from HDL2
by the liver and is converted back into HDL3
 Lipoproteins and Heart Disease
 LDL can deposit cholesterol in the arteries and
hence cause atherosclerosis which is the
underlying cause of heart attacks and stroke
 HDL can remove cholesterol from the artery wall
and thus prevent or even reverse atherosclerosis
 Thus the relative amount of LDL/HDL is an
important determinant of risk of developing
atherosclerosis
 What happens to fatty acids when
they get into cells?
Fatty Acids

Adipose Tissue Liver Muscle

Triacylglycerol Triacylglycerol Acetyl CoA Acetyl CoA

Phospholipid
CO2 Ketone bodies* CO2
* only in starvation
Fatty Acid Metabolism in Adipose Tissue
 Fatty Acid Metabolism in the Liver
VLDL

NEFA

Chylo
Remnant
secretion
lipid droplets
FA Esterification
(TAG & PL)

β-oxidation
(ketogenesis)
lipogenesis
Fatty Acid Metabolism in Muscle

Chylo/VLDL NEFA

FA

FA
-oxidation
Acetyl CoA
TCA Cycle

CO2 (+ATP)
 Fatty Acids As A Source of Energy
 Many tissues can oxidize fatty acids to produce
energy
– Muscle is quantitatively the most important
– Brain cannot utilize fatty acids but after prolonged
starvation it can use ketone bodies which are made
from fatty acids in the liver
 Three sources of fatty acid for energy
– Released by LPL from chylomicrons
– Released by LPL from VLDL
– Released as NEFA from adipose tissue
 Fatty Acid Oxidation
Linked to CoA before degradation

CH3(CH2)nCOOH + ATP + HS-CoA

Acyl CoA Synthase

CH3(CH2)nC-S-CoA + AMP + PPi

O
 Acyl-CoA has to move into the
mitochondria

Acyl-CoA CoA

Carnitine Acyl-Carnitine
mitochondria
Carnitine Acyl-Carnitine
Acyl-CoA CoA
cytosol
movement catalyzed by cartine palmitoyltransferase
Fatty Acid Oxidation
R-CH2-(CH2-CH2)-CO-S-CoA
FAD FADH2 oxidation
H
R-CH2-(C=C)-CO-S-CoA
H
H2O hydration

HO H
R-CH2-(C - C)-CO-S-CoA
H H
NAD NADH2 oxidation

O
R-CH2-(C -CH2)-CO-S-CoA
 Fatty Acid Oxidation cont.

O
R-CH2-C –(CH2-CO)-S-CoA
Thiolase
CoA-SH CH3-CO-S-CoA
Acetyl-CoA

R-CH2-CO-S-CoA
Acyl-CoA less
2 Carbons

The whole thing is then repeated removing 2 carbons each time

 Energy Yield from Palmitic Acid
(C16:0)
Overall Equation
Palmitoyl-CoA + 7FAD + 7NAD+ + 7CoA + 7H20
8Acetyl-CoA + 7FADH2 + 7NADH2
7 FADH2 7 x 1.5 = 10.5 ATP
7NADH2 7 x 2.5 = 17.5 ATP
8Acetyl-CoA 8 x 10 = 80 ATP
TOTAL = 108 ATP
However, when make Palmitoyl-CoA

ATP AMP = equivalent of 2ATP

Therefore, OVERALL TOTAL = 106

 Comparison of fatty acid synthesis & oxidation
 Site in cell
– Synthesis - cytosol
– Oxidation - mitochondria
 Carrier molecule
– Synthesis - ACP
– Oxidation - CoEnzyme A
 Enzymes
– Synthesis - Multifunctional Enzyme
– Oxidation - Separate enzymes
 Carbon Units
– Synthesis - 2C added from (3C) Malonyl- CoA
– Oxidiation - Yields (2C) Acetyl-CoA
 CoFactor
– Synthesis - Uses NADPH
– Oxidation - Yields NADH
 You should now be able to
describe
• The structure of fatty acids can differ (chain length, number,
position & type of double bond).
• The synthesis of fatty acids in animals (C16:0, C18:0 & C18:1).
• The synthesis of triacylglycerol and (glycero)phospholipids.
• The structure of lipoproteins and details of the 3 lipoprotein
pathways (exogenous, endogenous & reverse cholesterol
transport).
• The metabolism of fatty acids in different tissue (adipose tissue,
liver & muscle)
• -Oxidation of fatty acids