Metabolism of Lipids

Cholesterol Metabolism
AMU, School of Medicine
09/29/2020 Lipid Metabolism by Teklu 1

Biological significance of cholesterol
Notes: Contrary to popular belief, the biological role of cholesterol is not
limited to being the bad. Instead, it has a number of essential physiological
functions:
1. In humans and animals, cholesterol is a major constituent of the cell
membranes. Cholesterol modulates physical properties of these membranes
that in turn affect the function of membrane proteins such as receptors and
transporters. And also for posttranslational modification of membrane
proteins. Experimental depletion of membrane cholesterol cripples many
cellular functions.
2. Cholesterol is the biosynthetic precursor of bile acids, which are essential for
fat digestion.
3. Cholesterol is the precursor of all steroid hormones, namely, androgens,
estrogens, progestins, glucocorticoids, mineralocorticoids, and calciferol
(vitamin D).
Nevertheless, it certainly is true that cholesterol also plays a major role in the
pathogenesis of atherosclerosis.
Processes that determine the cholesterol balance
1. Intestinal uptake of dietary cholesterol
2. De novo cholesterol synthesis
3. Synthesis of steroid hormones from cholesterol
4. Synthesis of bile acids from cholesterol, and their biliary secretion.
5. Biliary secretion of surplus cholesterol in unmodified form.
Notes: Cholesterol can both be synthesized endogenously
(700mg/day) and obtained from the diet (about 400mg/day). Note that
significant amounts of cholesterol only occur in meat, eggs, and milk
products; plants and mushrooms contain other sterols but very little
cholesterol. The fact that vegetarians survive tells us that our capacity
to synthesize cholesterol suffices to cover our need for cholesterol
entirely.
However, we cannot degrade it; therefore, cholesterol has no
significant role in energy metabolism.
Role of Liver for cholesterol balance
 It is of critical importance that cells of the major tissues of the body be

assured a continuous supply of cholesterol.
 To meet this need a complex series of transport, biosynthetic and
regulatory mechanisms has evolved.
 The liver plays a central role in the regulation of the body’s cholesterol
balance.
1. Cholesterol enters the liver’s cholesterol pool from a number of sources
(influx) , including:
a. Dietary cholesterol.
b. Cholesterol synthesized by the extrahepatic tissues.
c. De novo synthesized cholesterol by the liver itself.
2. Cholesterol is eliminated (efflux) from the liver
a. As unmodified cholesterol in bile.
b. As a component of plasma lipoproteins sent to peripheral tissues in the form of
very low density lipoprotein (VLDL) Or
c. As bile salts secreted into the intestinal lumen.
Role of liver for cholesterol balance...
 The availability of cholesterol in the diets of different human
populations vary widely ; even in the same individual the amount
of cholesterol intake may change significantly from day to day.
 Therefore, regulatory mechanisms must exist to balance the rate of
cholesterol synthesis within the body against the rate of cholesterol
excretion.
 An imbalance in this regulation can lead to an elevation in
circulating levels of plasma cholesterol, causing the possibility of
coronary artery disease, whereas, excessive secretion of cholesterol
into the bile can result in precipitation of cholesterol in the gall
bladder and bile duct.

De-novo Synthesis of cholesterol
 Cholesterol is a sterol synthesized by virtually all tissues in
humans, although the largest contributions to the body’s
cholesterol are made by:
a. Liver
b. Intestine
c. Adrenal cortex
d. Reproductive tissues ( ovaries and testes)
e. Placenta
 Cholesterol synthesis occurs in the cytoplasm with enzymes in
both the cytosol and the endoplasmic reticulum.

A. Synthesis of HMG CoA
 The first two reactions in cholesterol synthetic pathway
are similar to those in the pathway that produces ketone
bodies; they result in the production of HMG CoA (β-
hydroxy - β- methylglutaryl CoA) except sub-cellular
location difference.
First, thiolase catalyzes the condensation of two acetyl

CoA molecules to form acetoacetyl CoA . Next, HMG
CoA synthase catalyzes the addition of a third molecule of
acetyl CoA producing HMG CoA.

A. Synthesis of HMG CoA…
 Liver parenchymal cells contain two isoenzyme forms of HMG
CoA synthase. The cytosolic enzyme participates in cholesterol
synthesis, whereas, the mitochondrial enzyme functions in the
pathway for ketone bodies synthesis.
 HMG-CoA destined for sterol synthesis is formed in the cytosol
and therefore, like the synthesis of fatty acids, cholesterol
biosynthesis depends on the export of acetyl-CoA from the
mitochondria.
 Also as with fatty acids, multiple steps in the cholesterol synthesis
require NADPH. The synthesis of cholesterol also requires ATP.

Synthesis of cytoplasmic HMG-
CoA
The acetyl CoA is provided by the
ATP-citrate lyase reaction as in the
case of fatty acid synthesis. Two
molecules of acetyl CoA condense
to form acetoacetyl CoA catalysed
by cytoplasmic acetoacetyl CoA
synthase

B. Synthesis of mevalonate from HMG-CoA
All steps downstream of HMG-CoA occur in the smooth

endoplasmic reticulum.
HMG-CoA reductase reduces HMG-CoA to mevalonate; uses 2
molecules of NADPH. This reaction is unique to the cholesterol
biosynthetic pathway. It is the rate-limiting step. Mevalonate is
converted to various isoprene intermediates.

C. Production of 5 Carbon Unit
Mevalonate is successively phosphorylated to phospho-mevalonate, to

pyrophosphomevalonate, then to 3-phospho-5 pyrophosphomevalonate. This then
undergoes decarboxylation to give isopentenyl pyrophosphate, a 5 carbon unit.
In the subsequent steps of the pathway, six molecules of isopentenyl-
pyrophosphate are used for the synthesis of one cholesterol molecule.

D. Condensation of 5-Carbon Units
Several rounds of “polymerization” of isoprenoid unit produce the
linear hydrocarbon molecule squalene, which is oxidized and cyclized
to the first sterol intermediate to be synthesized/lanosterol/. Squalene
undergoes oxidation by epoxidase, using molecular oxygen and
NADPH to form squalene epoxide. A cyclase converts it to 30C
lanosterol

F. Conversion of lanosterol to cholesterol
The 3 additional methyl groups on carbon atoms 4 and 14 are

removed to produce zymosterol.
Then the double bond migrates from 8-9 position to 5-6 position,
when desmosterol is formed. Desmosterol is present in fetal brain.
It is absent in adult brain and reappears in gliomas (brain tumor).
Finally, the double bond in the side chain (between carbon 24-25)
is reduced by NADPH when cholesterol is formed

Regulation of cholesterol synthesis
 HMG CoA reductase is an intrinsic membrane protein of the
endoplasmic reticulum; the enzyme’s active site extends into the cytosol.
HMG CoA reductase is the rate –limiting enzyme in cholesterol
synthesis, and is subject to different kinds of metabolic control:
1. Feed back inhibition: Cholesterol is a feed back inhibitor of HMG CoA
reductase, thus decreasing further cholesterol synthesis.
2. Hormonal regulation: (through phosphorylation/ dephosphorylation)=
short term regulation.
 Glucagon favors the formation of the inactive (phosphorylated) form of
HMG CoA reductase and hence, decreases the rate of cholesterol
synthesis.
 In contrast, insulin favors the formation of the active( dephosphorylated)
form of HMG CoA reductase and results in an increase in the rate of
cholesterol synthesis.

3. Sterol- mediated regulation of transcription (Gene regulation)= long term regulation
The synthesis of cholesterol is also regulated by the

amount of cholesterol taken up by the cells during
lipoprotein metabolism.
Chylomicron remnants internalized by liver cells , and
low density lipoproteins ( LDL ) internalized by the cells of
peripheral – tissues, provide cholesterol, which causes a
supress in transcription of the HMG CoA reductase gene,
leading to a decrease in de novo cholesterol synthesis.

The SREBP cleavage activator protein(SCAP), is an intracellular cholesterol sensor.
When cholesterol is less, SCAP escorts SREBP to Golgi bodies. Two Golgi proteases
(S1P and S2P) sequentially cleave the SREBP to a protein which binds to SRE and
activates transcription of HMG CoA reductase gene.

4. Inhibition by drugs:
The statin drugs (atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin,
and simvastatin) are structural analogs of HMG CoA, and are (or are metabolized
to) reversible, competitive inhibitors of HMG CoA reductase. They are used to
decrease plasma cholesterol levels in patients with hypercholesterolemia.

Bile acids and bile salts
 Bile: consists of a watery mixture of organic and inorganic

compounds.
 Organic components:
a. Lecithin (phosphatidylcholine)
b. Unmodified Cholesterol
c. Bile salts
 The “primary bile acids” are cholic acid and chenodeoxycholic acid.
Bile acids are amphipathic in that all the hydroxyl groups are α in
orientation and the methyl groups are β.Therefore, the molecules
have a polar and a non-polar face and can act as emulsifying agents
in the intestine, helping to prepare dietary TAG and other complex
lipids for degradation by pancreatic digestive enzymes.
 Bile salts provide the only significant mechanism for cholesterol
excretion, both as a metabolic product of cholesterol and as an
essential solubilizer for cholesterol excretion in bile.
Bile salt deficiency causes cholelithiasis

Synthesis of bile acids
Bile acids are synthesized in the liver. The

rate limiting step in bile acids synthesis is the
introduction of a hydroxyl group at carbon 7 of
the steroid ring by 7- α hydroxylase, which is
inhibited by cholic acid.

Synthesis of bile salts
Before the bile acids leave the liver,

they are conjugated to a molecule of
either glycine or taurine by an amide
bond between the carboxyl group of the
bile acid and the amino group on the
added compound.
These new structures are called bile

salts and include glycocholic acid and
glycochenodeoxycholic acids and
taurocholic acid and
taurochenodeoxycholic acids

Action of intestinal flora on bile salts
 Bacteria in the intestine can remove glycine and taurine from bile
salts, regenerating bile acids. They can also convert some of the
primary bile acids into “secondary” bile acids by removing a
hydroxyl group, producing deoxycholic acid from cholic acid and
lithocholic acid from chenodeoxycholic acid.
Enterohepatic circulation
 Bile salts secreted into the intestine are efficiently reabsorbed (greater
than 95%) and reused. The liver converts both primary and secondary
bile acids into bile salts by conjugation with glycine or taurine, and
secretes them into the bile. The mixture of bile acids and bile salts is
absorbed primarily in the ileum via a Na+-bile salt co-transporter.
 They are actively transported out of the ileal mucosal cells into the
portal blood, and are efficiently taken up by the hepatocytes via an
isoform of the co-transporter.

Enterohepatic circulation of bile salts and bile acids

Cholesteryl esters
 Much of the plasma cholesterol is in an

esterified form, with a fatty acid attached
at carbon – 3, which makes the structure
even more hydrophobic. Because of its
hydrophobicity, cholesterol must be
transported either in association with
protein as a component of a lipoprotein
particle or solubilized by phospholipid
and bile salts in bile.

Transport of cholesterol
 Cholesterol that is taken up from the intestines initially
becomes bound to chylomicrons.
 It may then be transferred from there to cells in the periphery
recall that chylomicrons bypass the liver or be redistributed
to other lipoprotein species. Some cholesterol will remain
with the chylomicron remnants that are finally phagocytosed
and degraded in the liver.
 The major site of cholesterol biosynthesis is once more the
liver. The major lipoprotein species synthesized by the liver
is known as very low density lipoprotein (VLDL), which
transports both cholesterol and triacylglycerol from the liver
to the periphery.
Transport of cholesterol…
 By extraction and exchange of triacylglycerol, the
composition of circulating VLDL successively changes until
it becomes low density lipoprotein (LDL), which has a
particularly high concentration of cholesterol.
 This lipoprotein species is removed from the circulation
through endocytosis by cholesterol-requiring cells, for
example in tissues that synthesize steroid hormones.
Endocytosis requires a specific receptor, the LDL receptor.
 A genetic deficiency of this receptor is associated with
increased blood levels of LDL and increased risk of heart
attack and stroke

Transport of cholesterol…
 Excess cholesterol can be transported back from peripheral
tissue to the liver by high density lipoprotein (HDL). Among all
lipoproteins, this one has the highest ratio of protein to lipid and
therefore the highest density.
 A high level of HDL in the blood is associated with a decreased
risk of cardiovascular disease; therefore, HDL has been dubbed
the ‘good cholesterol’ as opposed to LDL, the ‘bad cholesterol’.
 The liver can either recycle the cholesterol into new
lipoproteins, or dispose of excess cholesterol through secretion
into the bile.

Cholesterol in atherosclerosis
 Healthy arteries are flexible, strong and elastic. Over time, however,
too much pressure in your arteries can make the walls thick and stiff
— sometimes restricting blood flow to your organs and tissues.
This process is called arteriosclerosis, or hardening of the arteries.
 The damage may be caused by: High cholesterol, High blood
pressure, An irritant, such as nicotine, Certain diseases, such as
diabetes
 The level of plasma cholesterol, and in particular LDL-associated
cholesterol, is one of the main risk factors of atherosclerosis, the
degenerative vascular disease underlying myocardial infarction and
stroke. In western countries, this is the leading cause of death, being
more common than all cancers and leukemias combined.
 Briefly, atherosclerotic lesions develop as follows:

Development of atherosclerosis
A. High blood pressure promotes the formation of small defects in the
endothelium. These initial lesions allow blood plasma carrying LDL to seep
into the sub-endothelial tissue; this is followed by transmigration of
macrophages. Reactive oxygen species and enzymes released by
macrophages modify the LDL.
B. Modified LDL is taken up by macrophages. Lipid overload turns
macrophages into foam cells.
C. Foam cells perish, disintegrate and release the accumulated cholesterol,
which forms crystalline deposits. These crystals activate new macrophages
and cause them to release inflammatory cytokines that further incite and
amplify the inflammation
D. In an advanced lesion, cells in the muscular layer proliferate, progressively
constricting the artery. When the endothelium that covers the lesion becomes
eroded, thrombocytes and plasmatic coagulation factors are activated and
initiate blood clotting, causing acute thrombus formation and obstruction.

 As an alternative to thrombus formation, acute failure of
a damaged artery can also occur through rupture. About,
approximately 20% of all cerebral infarctions are caused
by rupture and hemorrhage rather than thrombotic
occlusion. Hemorrhage is less common in other organs.

How does LDL become oxidized?
 Transition metals (Fe, Cu) convert O2 to reactive oxygen

species: hydrogen peroxide (H2O2), superoxide (•O−O–)
 Macrophages produce reactive oxygen species
 Lipoxygenases produce lipid hydroperoxy-radicals that
can bind to LDL and induce lipid peroxidation
 Myeloperoxidase produces hypochlorite (HOCl)

Therapy of hypercholesterolemia
1. Limiting the dietary intake of cholesterol.

2. Vegetables are also rich in poly-unsaturated fatty acids (those with
several C.C double bonds) and in carbohydrate fibers, which seem to
have a greater favorable effect than restriction of cholesterol itself.
3. Inhibiting the intestinal uptake of cholesterol. Uptake can also be
inhibited by ezetimibe
4. Inhibition of endogenous cholesterol synthesis.
5. Promotion of cholesterol elimination, by way of bile acid depletion.
Ingested cholestyramine will bind bile acids and simply take them
along down the pipe. To replace the lost bile acids, the liver will
increase their synthesis and therefore deplete the pool of cholesterol.

Plasma lipoproteins
 Plasma lipoproteins are molecular complexes of lipids and
specific proteins.
 Plasma lipoproteins are dynamic particles which are in a
constant state of synthesis, degradation and removal from
the plasma.
Lipoprotein particles include:
a. Chylomicrons (CM)
b. Very low density lipoproteins (VLDL)
c. Low density lipoproteins (LDL)
d. High density lipoproteins (HDL)
Plasma lipoproteins…
 Lipoproteins function both to keep lipids soluble as they transport them in

plasma, and to provide an efficient mechanism for delivering their lipid contents
to the tissues.
 In humans the delivery system is less perfect than in other animals and as a result,
humans experience a gradual deposition of lipid , especially cholesterol in tissues.
 This is a potentially life-threatening occurrence when cholesterol deposition
contributes to plaque formation , causing narrowing of blood vessels, a condition
known as atherosclerosis .
 Lipoprotein particles constantly interchange lipids and apolipoproteins with each
other, therefore , the actual apolipoprotein and lipid content of each class of
particles can somewhat be variable.

Plasma lipoproteins…
 The principal lipids carried by lipoprotein particles are triacylglycerols and

cholesterol( free or esterified) obtained from diet or de-novo synthesis.
Composition of plasma lipoproteins
 Lipoproteins are composed of a neutral lipid core containing triacylglycerol
or cholesteryl esters or both, surrounded by a shell of apolipoproteins
(apoproteins), phospholipid and non-esterified cholesterol ( less
hydrophobic than esterified –cholesterol) , all oriented so that their polar
portions are exposed on the surface of the lipoprotein, thus making the
particle soluble in aqueous solution.

The lipoprotein particle. The external monolayer of a lipoprotein
particle contains free cholesterol, phospholipids, and apoproteins.
Cholesterol esters and triacylglycerols locate in the particle core.
Approximate size& density
of serum lipoproteins

Table: Plasma lipid profile (normal values)

Apolipoproteins (Apoproteins)
 Apolipoproteins are embedded into the surface of the particle. These
protein molecules mainly serve as “address tags” that mediate the
interaction with target molecules and cells, such as lipoprotein lipase
on endothelial cells or the LDL receptor.
 Some apolipoproteins are stably associated with their lipoprotein
particles; for example, a given molecule of apolipoprotein B remains
associated with the same chylomicron or VLDL particle throughout
its lifetime.
 Exchangeable apolipoproteins have regulatory roles; for example,
apolipoprotein CII reversibly associates with VLDL particles and
activates their interaction with lipoprotein lipase.
 Apolipoproteins are divided by structure and function into classes A
to H, with most classes having sub-classes , for example, apo A- I
and apo C- II.
Characteristics of apoproteins and their functions

Metabolism of chylomicrons
 Apolipoprotein synthesis begins on the rough endoplasmic reticulum
apolipoproteins are glycosylated as they move through the ER and
Golgi.
 The enzymes involved in TAG, cholesterol and phospholipid
synthesis are located in the smooth ER.
 Assembly of the apolipoproteins and lipid into chylomicrons occurs
during transition from the ER to Golgi, where they are packaged in
secretory vesicles that are exported from the cell into the lymphatic
system by the lacteals.

 When chylomicron reaches the plasma, the nacent chylomicron is
rapidly modified, receiving apoE which in conjunction with apoB-
48, is recognized by hepatic receptors and C
apolipoproteins( including apo C- II, necessary for activation of
lipoprotein lipase, the enzyme that degrades the TAG contained in
the chylomicron).
 The source of these apolipoproteins is circulating HDL.
 Lipoprotein lipase is found predominantly in the
capillaries of the adipose tissue and cardiac and skeletal
muscle.

 Chylomicron remnants bind to apoB-48 / apo E receptors on
hepatocyte membranes and are taken into the cells by endocytosis.
 The endocytozed vesicle then fuses with a lysosome and the
apolipoproteins, cholesteryl esters and other components of the
remnant are hydrolytically degraded, releasing amino acids, free
cholesterol and fatty acids.
 The cholesterol released from the chylomicron regulates the rate
of the de novo cholesterol synthesis in the liver by causing a
decrease in cell content of HMG CoA reductase as well as by
allosterically inhibiting the enzyme.

Metabolism of very low density lipoproteins
 VLDLs are produced in the liver. They are composed
predominantly of TAG, and their function is to carry this lipid
from the liver to peripheral tissues.
 There, the TAG is degraded by lipoprotein lipase (as
Chylomicrons)
 Remember Fatty liver occurs in conditions in which there is an
imbalance between hepatic TAG synthesis and the secretion of
VLDL. Diseases such as hepatitis, uncontrolled diabetes mellitus
and chronic ethanol ingestion can cause fatty liver.

 Abetalipoproteinemia, which is due to a lack of MTP (microsomal
triglyceride transfer protein) activity, results in an inability to assemble
both chylomicrons in the intestine and VLDL particles in the liver.
 The triacylglycerol, which is produced in the smooth endoplasmic
reticulum of the liver, is packaged with cholesterol, phospholipids, and
proteins (synthesized in the rough endoplasmic reticulum) to form
VLDL .
 The microsomal triglyceride transfer protein (MTP), which is required
for chylomicron assembly, is also required for VLDL assembly. The
major protein of VLDL is apoB-100.
 There is one long apoB-100 molecule wound through the surface of
each VLDL particle.
 Similar to chylomicrons, VLDL particles are first synthesized in a
nascent form, and on entering the circulation they acquire apoproteins
CII and E from HDL particles to become mature VLDL particles.

Synthesis of VLDL from Glucose

Modification of circulating VLDL
 As VLDLs pass through the circulation their structure is altered, TAG is

removed by lipoprotein lipase, causing the VLDL to decrease in size and
become more dense.
 Surface components, including the apoC-II and apo E apolipoproteins ( that
originally had been donated to the VLDL from HDL), are transferred to HDL.
 Finally, cholesteryl esters are transferred from HDL to VLDL in an exchange
reaction that concomitantly transfers TAG or phospholipid from VLDL to
HDL.
 This exchange is accomplished by cholesteryl ester transfer protein.

Production of LDL from VLDL in plasma
 After these modifications, the VLDL has been converted in
the plasma to LDL.
 An intermediate- sized particle, the intermediate density
lipoprotein ( IDL) is observed during transition from VLDL to
LDL in the plasma.
 IDLs can also be taken up by cells through receptor-mediated
endocytosis.

Metabolism of low density lipoproteins
 LDL particles retain apoB – 100 , but lose their other aplipoproteins to
HDL. They contain much less TAG than their VLDL predecessors, and
have a high concentration of cholesterol and cholesteryl esters.
1. Receptor- mediated endocytosis:
 The primary function of LDL particles is to provide cholesterol to

peripheral tissues.
 They do so both by depositing free cholesterol on the membranes of cells
as they come in contact with the cell surface, and by binding to receptors
on the cell- surface membranes that recognize apolipoprotein B-100.
 A similar mechanism is used for the cellular uptake and degradation of
chylomicron remnants and HDLs by the liver.
 LDL receptors are negatively charged glycoprotein molecules. A deficiency of
functional LDL receptors causes a significant elevation in plasma LDL, and
therefore of plasma cholesterol, but plasma TAG levels remain normal, for
example, as in type II hyperlipidemia (familial –hyperbeta- lipoproteinemia). This
can greatly accelerate the progress of atherosclerosis.
2. After binding, the LDLs are internalized as intact particles by endocytosis.
3. The vesicle containing the LDL rapidly loses its clathrin coat and fuses with other
similar vesicles, forming larger vesicles called endosomes.
4. The pH of the contents of the endosome falls ( due to the proton-pumping activity
of endosomal ATP ase), allowing the separation of the LDL from its receptor. The
receptors then migrate to one side of the endosome, whereas, the LDLs stay free
within the lumen of the vesicle.

 The receptors can be recycled, whereas, the lipoprotein remnants in the vesicle are
degraded by lysosomal ( hydrolytic) enzymes, releasing cholesterol, amino acids,
fatty acids and phospholipids. =These compounds can also be recycled by the cell.
 The number of receptors for lipoproteins(LDL –receptors) varies according to the
availability of these lipoprotein particles and according to the needs of the cell.
 The LDL (apo B-100, E) receptor is a “high-affinity” LDL receptor, which may be
saturated under most circumstances.
 For example, if there is a large amount of a particular circulating plasma
lipoprotein (LDL), the number of cell- surface receptors for it decreases, frequently
termed “ down –regulation”. Conversely, if cells are starved for cholesterol, they
increase the number of cell- surface receptors “ up- regulation”.

Effect of endocytosed cholesterol on cell cholesterol content
 The chylomicron remnant, HDL and LDL –derived cholesterol affects cellular
cholesterol content in several ways:
1. HMG CoA reductase activity is inhibited by cholesterol so that de novo
cholesterol synthesis decreases.
2. If the cholesterol is not required immediately for some structural or synthetic
purpose, it is esterified by acyl CoA : cholesterolacyltransferase (ACAT).
ACAT transfers a fatty acid from a fatty acyl CoA derivative to cholesterol,
producing a cholesteryl ester that can be stored in the cell. The activity of this
enzyme is enhanced in the presence of increased intracellular cholesterol.
3. Synthesis of new LDL –receptor protein is lowered by decreasing
transcription of the LDL-receptor gene.
Metabolism of high-density lipoprotein (HDL)
HDL particles are synthesized in the liver and released into the blood
stream by exocytosis. They perform a number of important
functions:
1. Serve as a circulating reservoir of apo C-II, the apolipoprotein that
is transferred to VLDL and chylomicrons and is an activator of
lipoprotein lipase. Also it acts as a reservoir for apo –E, necessary
for the recognition of chylomicron remnants by the liver receptors.

2. Removes free (unesterified) cholesterol from extrahepatic tissues and esterifying
it , using the plasma enzyme phosphatidylcholine : cholesterol acyltransferase,
(PCAT ) also known as LCAT, where” L “stands for lecithin). LDL is taken up by
cells in the periphery through endocytosis, which is mediated by the LDL receptor.
3. Excess cholesterol is exported from the cell by an active transporter (ABCA1) and
delivered to high density lipoprotein (HDL), which then carries it back to the liver.
Cholesterol transport by HDL is facilitated by lecithin-cholesterol acyltransferase.
4. Transfers cholesteryl to VLDL and LDL in exchange for TAG.
5.Carries cholesteryl esters to the liver, where the HDL is degraded and cholesterol
released.

HDL uptake of free cholesterol
 Newly secreted HDLs are disc shaped particles containing predominantly
unesterified cholesterol, phospholipid (largely phosphatidylcholine) , and a
number of apolipoproteins including apoE, apoA-I and apoC-II.
 They are rapidly converted to spherical particles as they accumulate
cholesterol.
 HDL particles are excellent acceptors of unesterified cholesterol from the
surface of cell membranes and from other circulating lipoproteins.
 As the nacent HDL accumulates cholesteryl esters it first becomes
classified as HDL3 and eventually becomes a round, micellar –like particle
HDL 2.

Esterification of free cholesterol in HDL
 Once free cholesterol is taken up by the HDL particle, it is
immediately esterified by PCAT, a plasma enzyme synthesized by the
liver, which is activated by apoA-1 of the HDL. Plasma levels of
apoA-1 are increased by modest alcohol intake.
 Notes: The HDL particle contains the enzyme lecithin cholesterol
acyltransferase, or LCAT for short, which converts cholesterol to
cholesterol esters. The LCAT reaction occurs at the surface of HDL
particles. The transfer of one acyl chain from a lecithin
(phosphatidylcholine) molecule to cholesterol produces a cholesterol
ester and lysolecithin.
Esterification of free cholesterol in HDL…
 Notes: Cholesterol is amphiphilic and tends to accumulate at
lipid/water interfaces, with the OH group exposed to the aqueous
phase. Free cholesterol can therefore be transported only within the
outermost layer of lipid molecules of a lipoprotein particle.
 In contrast, cholesterol esters are entirely hydrophobic and readily
partition into the interior of lipoprotein particles. The LCAT reaction
therefore greatly increases the transport capacity of HDL particles for
cholesterol.
 The resulting cholesteryl ester is so hydrophobic that is effectively
trapped in the HDL and can no longer be transferred to a membrane.
 The only mechanism for removing it from HDL in the plasma is
through transfer to VLDL by cholesterol ester transfer protein, where it
ultimately remains in the LDL until the particle is taken up by a cell.
 About 2/3 of the cholesterol in the plasma is esterified with fatty acid.
In liver disease, a decreased concentration of plasma cholesteryl esters
is observed.
 This may be due to either a deficiency in phosphatidylcholine
production or a lack of PCAT.

Fate of HDLs
 Spherical HDL particles are taken up by the liver by receptor-
mediated endocytosis, and cholesteryl esters are degraded.
 The cholesterol thus released can be either repackaged in
lipoproteins, converted into bile acids or secreted into the bile
for removal from the body.

Reverse cholesterol transport
 The selective transfer of cholesterol from peripheral cells to HDLs and from HDLs
to the liver for bile acids synthesis or disposal via the bile and to steroidogenic cells
for hormone synthesis is a key component of cholesterol homeostasis.
 This is in part the basis for the inverse relationship seen between plasma HDL
concentration and atherosclerosis and for HDL’s designation as the “ good
cholesterol carrier”.
 Reverse cholesterol transport involves efflux of cholesterol from peripheral cells to
HDL, esterification of cholesterol by PCAT, binding of the cholesterol ester-rich
HDL (HDL2) to the liver and steroidogenic cells, the selective transfer of
cholesteryl esters into these cells and the release of lipid-depleted HDL (HDL3).
 The process is thought to be mediated by a cell-surface receptor (scavenger
receptor class B, SR-B 4) that binds HDL.

Role of lipoprotein (a) in heart disease
 Lipoprotein (a) is a particle that , when present in large quantities in the

plasma, has been suggested to increase the risk of coronary heart disease.
 Lipoprotein (a) is nearly identical in structure to LDL particles; its one
distinguishing feature is the presence of an additional apolipoprotein
molecule, apolipoprotein(a), that is covalently linked at a single site to
apoB-100.
 80% of the amino acids in apolipoprotein(a) are the same as those of
plasminogen, the precursor of a blood protease whose target is fibrin, the
main protein component of blood clots.
 It is hypothesized that lipoprotein (a) may slow down the breakdown of
blood clots that trigger heart attacks and cause cell proliferation. This is
because it competes with plasminogen for the binding of plasminogen
activators.

Metabolism of the Eicosanoids
 “Eicosa” is the Greek word for the number 20. Eicosanoids are
derived from polyunsaturated fatty acids containing 20 carbon
atoms, which are found in cell membranes esterified to membrane
phospholipids.
 Arachidonic acid, derived from the diet or synthesized from
essential fatty acid , linoleic acid. It is elongated and desaturated
to arachidonic acid, the immediate precursor of the predominant
class of prostaglandins (those with two double bonds) in humans.
 Arachidonic acid, is released from membrane- bound
phospholipids by phospholipaseA2 in response to a variety of
signals ( angiotensin II, bradykinin, epinephrine and vasopressin).

Release of arachidonic acid from membrane

Metabolism of the Eicosanoids…
 Arachidonic acid is enzymatically metabolized by three major
pathways.
 Three major pathways for the metabolism of arachidonic acid occur
in various tissues.
1. The first of these, the cyclooxygenase pathway, leads to the
synthesis of prostaglandins and thromboxanes.
2. The second, the lipoxygenase pathway, yields the leukotrienes,
HETEs, and lipoxins.
3. The third pathway, catalyzed by the cytochrome P450 system, is
responsible for the synthesis of the epoxides, HETEs, and diHETEs.
 Many eicosanoids have very short half-lives, in the range of a few
minutes or less. They are rapidly inactivated and excreted.

General structure of
Prostaglandins General structure of TXA2

Metabolism of the Eicosanoids…
 After arachidonic acid is released into the cytosol, it is
converted to eicosanoids by a variety of enzymes with
activities that vary among tissues.
 This variation explains why some cells, such as those in the
vascular endothelium, synthesize prostaglandins E2 and I2
(PGE2 and PGI2) whereas cells, such as platelets, synthesize
primarily thromboxane A2 (TXA2) and 12-
hydroxyeicosatetraenoic acid (12-HETE).
 Eicosanoids are classified in to two groups
1. Prostanoids : that include prostagladins, prostacyclins and
thromboxanes
2. Leukotrienes and lipoxins
Synthesis of PGH2
 The first step in the synthesis of prostaglandins is the oxidative
cyclization of the free arachidonic acid to yield PGH 2 by
prostaglandin endoperoxide synthase.
 This is a microsomal enzyme, that has two catalytic activities, fatty
acid cyclooxygenase (COX), which requires two molecules of O2
and peroxidase which is dependent on reduced glutathione.
 PGH2 is converted to a variety of prostaglandins and thromboxanes,
by cell-specific synthases.
 PGH2 is in turn metabolized to the prostanoid lipid signals (PGD2,
PGE2, PGF2α, PGH2, PGI2, or TXA2) by one of the secondary
enzymes that are named for the individual prostanoid produced. The
type of prostanoid produced is determined by which downstream
enzyme is present; usually one downstream enzyme predominates in
a given cell.
Isoenzymes of prostaglandin endoperoxide synthase:
 Two enzymes denoted as COX-1 and COX-2 of the synthase

are known.
 COX-1 is made constitutively in most tissues and is required
for maintenance of healthy gastric tissue, renal homeostasis
and platelet aggregation.
 COX-2 is inducible in a limited number of tissues in response
to products of activated immune and inflammatory cells. The
increase in prostaglandin synthesis subsequent to the induction
of COX-2, mediates the pain, heat redness, and swelling of
inflammation and fever of infection.

Inhibition of prostaglandin synthesis
 Synthesis of prostaglandins can be inhibited by a number of unrelated

compounds. For example, cortisol ( a steroidal anti-inflammatory agent
inhibits phospholipase A2 activity and therefore, the precursor of the
prostaglandins, arachidonic acid, is not available.
 Cortisol, also inhibits COX-2 , but not COX-1.
 Aspirin, indomethacin and phenylbutazone ( all non-steroidal anti-
inflammatory agents or NSAIDS) inhibit both COX-1 and COX-2 and,
therefore, prevent the synthesis of the parent prostaglandin , PGH2.
Systemic inhibition of COX-1 with subsequent damage to the stomach
and the kidneys and impaired clotting of blood, is the basis of aspirin
toxicity.
 Inhibitors specific for COX-2 e.g., celecoxib are designed to reduce
pathologic inflammatory processes while maintaining the physiologic
functions of COX-1.

Inhibition of prostaglandin synthesis…
 Inflammation involving the mucosal and smooth muscle layers of the

respiratory tract plays a major role in the development of acute asthmatic
bronchospasm.
 Dexamethasone and other potent glucocorticoids are capable of preventing
or suppressing this inflammation. In part, the glucocorticoids act by inhibiting
the recruitment of leukocytes and monocytes–macrophages into affected areas.
 They also limit the ability of these cells to elaborate a variety of chemotactic
factors and other substances, such as certain eicosanoids, which enhance the
inflammatory process.
 Glucocorticoids, for example, suppress the transcription and translation of the
inducible form of the cyclooxygenase enzyme, COX-2. Glucocorticoids also
induce the synthesis of a protein or family of proteins (lipocortins or
macrocortins) that inhibit the activity of phospholipase A2. As a result, the
synthesis of prostaglandins and leukotrienes is decreased, and the
inflammatory response in bronchial tissues is reduced

Inhibition of prostaglandin synthesis

The role of prostaglandins in platelet homeostasis
 Thromboxane A2 (TXA2), is produced by activated platelets. It promotes
adherence and aggregation of circulating platelets, and contraction of
vascular smooth muscle, thus promoting formation of blood clots (thrombi).
 Prostacyclin (PGI2), produced by vascular endothelial cells, inhibits
platelet aggregation and stimulates vasodilatation, and so impedes
thrombogenesis. The opposing effects of TXA2 and PGI2 limit thrombi
formation to sites of vascular injury.
 Aspirin has an antithrombogenic effect. It inhibits TXA2 synthesis from
arachidonic acid in platelets, by irreversible acetylation and inhibition of
COX-1. This irreversible inhibition of COX-1 cannot be overcome in
anucleate platelets, but can be overcome in endothelial cells, because they
have a nucleus and therefore can generate more of the enzyme.
 This difference is the basis of low -dose aspirin therapy used to lower the
risk of stroke and heart attacks by decreasing formation of thrombi.

Characteristic features of prostaglandins
1. Act as local hormones.

2. Show the effects near the site of synthesis(autocrine and
Paracrine effects)
3. Are not stored in the body
4. Have a very short life span and are destroyed within seconds or
few minutes
5. Production increases or decreases in response to diverse stimuli
or drugs
6. Are very potent in action. Even in minute (nanogram
concentration), biological effects are observed.

Synthesis of leukotrienes
 Leukotrienes were so named because they are synthesized in
leukocytes and contain the typical triene structure, i.e., three double
bonds in series (in this case, at positions 7, 9, and 11).
 Arachidonic acid is converted to a variety of linear hydroperoxy
acids by a separate pathway involving a family of lipoxygenases.
 For, example neutrophils contain 5-lipoxygenase , which
converts arachidonic acid to 5-hydroxy-6,8,11,14 eicosatetraenoic
acid (5-HPETE).
 5-HPETE is converted to a series of leukotrienes, the nature of the
final products varying according to the tissue. Lipoxygenases are
not affected by NSAIDS.
 Leukotrienes are mediators of allergic response and
inflammation. Inhibitors of 5-lipoxygenase and leukotriene
receptor antagonists are used in the treatment of asthma.
Action of lipoxygenases in the formation of HPETEs and HETEs.
Lipoxygenases add hydroperoxy groups at position 5, 12, or 15
with rearrangement of the double bond. HPETEs are unstable and
are rapidly reduced to form HETEs or converted to leukotrienes and
lipoxins

Metabolism of Lipids

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Metabolism of Lipids

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Cholesterol Metabolism

AMU, School of Medicine

09/29/2020 Lipid Metabolism by Teklu 1

 It is of critical importance that cells of the major tissues of the body be

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First, thiolase catalyzes the condensation of two acetyl

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All steps downstream of HMG-CoA occur in the smooth

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Mevalonate is successively phosphorylated to phospho-mevalonate, to

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The 3 additional methyl groups on carbon atoms 4 and 14 are

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The synthesis of cholesterol is also regulated by the

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 Bile: consists of a watery mixture of organic and inorganic

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Bile acids are synthesized in the liver. The

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Before the bile acids leave the liver,

These new structures are called bile

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 Much of the plasma cholesterol is in an

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 Transition metals (Fe, Cu) convert O2 to reactive oxygen

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1. Limiting the dietary intake of cholesterol.

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 Lipoproteins function both to keep lipids soluble as they transport them in

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 The principal lipids carried by lipoprotein particles are triacylglycerols and

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 As VLDLs pass through the circulation their structure is altered, TAG is

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1. Receptor- mediated endocytosis:

 The primary function of LDL particles is to provide cholesterol to

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