CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF.

HARPERS)

REVIEW QUESTIONS QUICK GUIDE

QUESTION ANSWER
BIOMEDICAL IMPORTANCE
Cholesterol is present in tissues and plasma as Free cholesterol
Combined with long chain FA as Cholesteryl ester
(storage form)
In plasma, both are transported as lipoproteins
Essential structural component of membranes Amphipathic lipid in cholesterol
Maintenance of permeability and fluidity
Outer layer of plasma lipoproteins
Synthesized from acetyl-CoA Cholesterol
Precursor of all steroids
Product of animal catabolism
Occurs in foods of animal origin (egg yolk, meat, liver,
brain)
Major constituent of gallstones
Examples of steroids Corticosteroids
Sex hormones
Bile acids
Vitamin D
Vehicle that supplies cholesterol and Cholesteryl ester to LDL
tissues
Removed from tissues by HDL Free Cholesterol
Transported to the liver
Eliminated either unchanged or after conversion to bile
acids
(Reverse cholesterol transport)
Chief role in pathologic process Atherosclerosis of vital arteries
-cerebrovascular
-coronary and peripheral vascular disease
CHOLESTEROL IS BIOSYNTHESIZED FROM ACETYL-COA
Sources of cholesterol More than half is synthesized by the body (700
mg/d)
Average diet
Account for approximately 10% each of total synthesis in Liver and intestine
humans
All tissues containing nucleated cells are capable of True
cholesterol synthesis. True or False?
Site of cholesterol synthesis ER and Cytoplasmic compartments
Source of all carbon atoms in cholesterol Acetyl-coA
Cholesterol is a 28 carbon compound consisting of 3 rings False.
and a side chain. True or false? 27 carbons
4 rings

Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

Synthesized from acetyl coa by a pathway divided into 5 1. synthesis of mevalonate from acetyl-coa
steps 2. formation of isoprenoid from mevalonate by
loss of co2
3. condensation of six isoprenoid units to form
squalene
4. cyclization of squalene give rise to Ianosterol
(parent steroid)
5. formation of cholesterol from Ianosterol
STEP 1 – BIOSYNTHESIS OF MEVALONATE
Formed by the reactions used in mitochondira to HMG-CoA
synthesize ketone bodies (3-hydroxy-3-methyglutaryl-CoA)
Cholesterol synthesis is extramitochondiral thus two True
pathways are distinct. True or False?
Catalyze the formation of acetoacetyl-CoA from two mols Thiolase
of acetyl-CoA
Catalyze the formation of HMG-CoA as acetoacetyl-CoA HMG-CoA Synthase
condenses with acetyl-CoA
Catalyze the reduction of HMG-CoA to mevalonate by HMG-CoA Reductase
NADPH
Principal regulatory step in the pathway of cholesterol Last step (reduction to mevalonate by NADPH
synthesis catalyzed by HMG-CoA Reductase)
Site of action of the most effective class of cholesterol
lowering drugs
A cholesterol lowing drug Statins
HMG-CoA Reductase inhibitor
STEP 2 – FORMATION OF ISOPRENOID UNITS
Phosphorylated using ATP by three kinases Mevalonate
Active isoprenoid unit formed after decarboxylation Isopentenyl diphosphate
STEP 3 – SIX ISOPRENOID UNITS FORM SQUUALENE
Isomerized by a shift of the double bond to form Isopentenyl diphosphate
dimethyallyl diphosphate
Isopentenyl diphosphate condensed with another Geranyl diphosphate
molecule of isopentenyl diphosphate to form the 10
carbon intermediate which is
Formed from further condensation with isopentenyl Farnesyl diphosphate
diphosphate
Formed from two molecules of farnesyl diphosphate Squalene
condense at the diphosphated end
Formed from elimination of inorganic pyrophosphate Presqualene diphosphate
Reduced by NADPG with elimination of a further inorganic
pyrophosphate molecule
STEP 4 – FORMATION OF LANOSTEROL
Squalene can fold and resemble a steroid nucleus. True or True
False?
Catalyze by a mixed function oxidase in the ER to convert Squalene epoxidase
squalene to squalene 2,3-epoxide before ring closure
occurs

Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

Catalyzed the transfer of methyl group on C14 to C13 and Oxidosqualene-Ianosterol cyclase
that on C8 to C14 as cyclization occurs
STEP 5 – FORMATION OF CHOLESTEROL
Site of cholesterol formation from Ianosterol Membranes of ER
Changes in the cholesterol formation from Ianosterol Steroid nucleus and side chain
Formed from removing C14 and C4 14-desmethyl Ianosterol
After the formation of 14-desmethyl Ianosterol Zymosterol
Formed from moving the double bond at C8’C9 to C5’C6 in Desmosterol
two steps
Formed from reducing the double bond of the side chain Cholesterol
FARNESYL DIPHOSPHATE GIVES RISE TO DOLICHOL AND UBIQUINONE
Forms the dolichol and ubiquinone by further addition of Farnesyl diphosphate
up to 16 (dolichol) or 3-7 (ubiquinone) isopentenyl
diphosphate residues
Some GTP—binding proteins in the cell membrane are Farnesyl geranylgeranyl (20 carbon) residues
prenalated with
Facilitate the anchoring of proteins into lipoid membranes Protein prenylation
Involved in protein-protein interaction and membrane
associated protein trafficking
CHOLESTEROL SYNTHESIS IS CONTROLLED BY REGULATION OF HMG-COA REDUCTASE
Regulated the cholesterol synthesis near the beginning of HMG-CoA reducatse
the pathway
Decrease synthesis of cholesterol in starving animals is False. Decrease activity of HMG-CoA Reductase
accompanied by increased activity of HMG-CoA
Reductase. True or False?
Only inhibited by dietary cholesterol Hepatic synthesis
Inhibit HMG-CoA in liver Mevalonate
Cholesterol
Immediate product of the reaction Mevalonate
Main product of the pathway Cholesterol
Repress transcription of the HMG CoA Reductase via Cholesterol and metabolites
activation of SREBP (sterol regulatory element binding
protein) transcription factor
Family of proteins involved in transcription of genes SREBP trans factor
involved in cellular uptake and metabolism of cholesterol
and other lipids
Inhibit SREBP activation Insig
Expression is induced by insulin (insulin induced gene)
Present in the ER
Promotes degradation of HMG-CoA Reductase
A diurnal variation occurs both in cholesterol synthesis True
and Reductase activity. True or False?
By regulating the rate of protein synthesis degradation, Posttranslational modification
enzyme activity is also modulated more rapidly by
A mechanism that reversibly modify activity Phosphorylation-dephosphorylation mechanisms
Some may be cAMP-dependent
Or Immediately responsive to glucagon
Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

Phosphorylates and inactivates HMG-CoA Reductase AMP-activated protein kinase (AMPK)

Activate via phosphorylation by AMPK kinase (AMPKK) an
allosteric modification by AMP
Increases HMG-CoA Reductase activity Insulin or Thyroid Hormone
Decrease HMG-CoA Reductase activity Glucagon or glucocorticoids
Lowering plasma cholesterol in humans by lowering True
amount of dietary cholesterol produce variable results.
True or false?
200 mg decrease of dietary cholesterol causes approx. False. 100 mg
0.13 mmol/L of serum decrease. True or False?
FACTORS THAT INFLUENCE THE CHOLESTEROL BALANCE IN TISSUES
Causes of increased cholesterol Uptake of cholesterol-containing lipoproteins
Uptake of free cholesterol from cholesterol rich
lipoproteins to the cell membrane
Cholesterol synthesis
Hydrolysis of Cholesteryl esters by the enzyme
Cholesteryl ester hydrolase
Causes of decreased cholesterol Efflux of cholesterol from membrane HDL via
ABCA1, ABCG1 or SR-B1
Esterification of cholesterol by ACAT
(acyl-CoA: cholesterol acyltransferase)
Utilization of cholesterol by synthesis of other
steroids such as hormones or bile acids in the liver
MANY FACTORS INFLUENCE THE CHOLESTEROL BALANCE IN TISSUES
Occur on cell surface in pits that are coated on the LDL (apo B-100, E) receptor
cytosolic side of the cell membrane with a protein called
clathrin
Spans the membrane, the B-100 binding region being at Glycogen receptor
the exposed amino terminal end
After binding, LDl is taken up intact by endocytosis
Site of apoprotein and Cholesteryl ester hydrolysis Lysosomes
After hydrolysis, cholesterol is translocated into the cell True
and receptors are recycled to the cell surface True or
False?
Transcription of the genes encoding enzymes inhibited by HMG-CoA synthase
influx of cholesterol HMG-CoA Reductase
Other enzymes involved in cholesterol synthesis
Coordinately suppresses cholesterol synthesis and uptake LDL receptor
via the SREBP pathway
When stimulated, promote cholesterol esterification ACAT activity
Regulates the recycling of the receptor to the cell surface Protein proprotein covertase subtilisin/kexin type
by targeting it for degradation 9 (PCKS9)
Regulate the LDL receptor activity on the cell surface Cholesterol requirement for membranes
Steroid hormones
Bile acid synthesis
Free cholesterol content of the cell is kept within relatively False. Narrow limits
high limits. True or false?

Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

CHOLESTEROL IS TRANSPORTED BETWEEN TISSUES IN PLASMA LIPOPROTEINS

Derived from intestinal absorption of TAG and other lipids Chylomicrons
Derived from the liver for the export of TAGS to VLDL
extrahepatic tissues
Representing a final stage in the catabolism of VLDL LDL
Involved in cholesterol transport and in VLDL and HDL
chylomicron metabolism
Greater part of a form of cholesterol transported in Cholesteryl esters
plasma lipoproteins
Highest proportion of Cholesteryl esters is found in LDL
Dietary cholesterol equilibrates with plasma cholesterol in False. Days
weeks. True or false?
Dietary cholesterol equilibrates with tissue cholesterol in False. Weeks
days. True or false?
Hydrolyzed to cholesterol, absorbed by the intestine Cholesteryl ester
together with dietary unesterified cholesterol and other
lipids
With cholesterol synthesized in the intestines, it is then True
incorporated into chylomicrons. True or False?
Of the cholesterol absorbed, 70-80% is esterified with long False. 80-90%
chain fatty acids in the intestinal mucosa. True or False?
95% of chylomicron cholesterol delivered to the liver Chylomicron remnants
Most of the cholesterol in VLDL is retained during the False. LDL not HDL
formation of intermediate-density lipoprotein and HDL
taken up by liver and extrahepatic tissues. True or False?
PLASMA LCAT IS RESPONSIBLE FOR VIRTUALLY ALL PLASMA CHOLESTERYL ESTER IN HUMANS
Associated with HDL + apo A-I LCAT
Draws in cholesterol from tissues and other lipoproteins HDL
once it is esterified
Enable it to function in reverse cholesterol transport
CHOLESTERYL ESTER TRANSFER PROTEIN (CETP) FACILITATES TRANSFER OF CHOLESTERYL ESTER FROM HDL TO
OTHER LIPOPROTEINS
Associated with HDL CETP
Found in human plasma (Cholesteryl ester transfer protein)
Facilitate transfer of Cholesteryl ester from HDL to VLDL,
IDL and LDL in exchange of TAGS relieving product
inhibition of the LCAT activity in HDL
Cholesteryl ester formed by LCAT finds its way to the liver VLDL remnants (IDL) or LDL
via
Delivers its cholesterol to the liver in HDL cycle TAGS-enriched HDL2
CHOLESTEROL IS EXCRETED FROM THE BODY IN THE BILE AS CHOLESTEROL OR BILE ACIDS (SALTS)
Cholesterol is excreted from the body via bile either in Unesterified form
After conversion into bile acids in the liver
Principal sterol in the feces Coprostanol
Formed from cholesterol by the bacteria in the lower
intestine
Synthesized in the liver form cholesterol Primary bile acids
Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

2 Primary bile acids Cholic acid (largest amount in most mammals)

Chenodeoxycholic acid
First and principal regulatory step in the biosynthesis of 7alpha-hydroxylation of cholesterol
bile acids
Catalyze the regulatory step in the biosynthesis of bile Cholesterol 7’-hydroxylase
acids
Microsomal cytochrome P450 enzyme designated CYP7A1
A typical monooxygenase
Requires O2
NADPH
Cytochrome P450
Catalyzes subsequent hydroxylation steps Monooxygenases
Pathway of bile acid biosynthesis divides early into CholylCoA (extra alpha-OG group on position 12)
subpathway leading to Chenodeoxycholyl-CoA
First step responsible for a significant proportion of the 27-hydroxylation of cholesterol by cytochrome
primary bile acids synthesized P450 sterol 27-hydroxylase (CYP27A1)
Primary bile acids enter the bile as Glycine or taurine conjugates
Where conjugates takes place Liver peroxisomes
Ratio of glycine to taurine conjugates 3:1
pH in which bile acids and their conjugates are assumed to pH 7.6-8.4
be in a salt form hence the term bile salts
Primary bile acids are further metabolized in the intestine
by intestinal bacteria then leads to deconjugation and
formation of 7alpha-dehydroxylation then formation of
secondary bile acids (deoxycholic acid and lithocholic acid)
takes place
Most bile acids goes out from the liver. True or False? False. It is returned to the liver in the
enterohepatic circulation
st
Site of absorption of fat including cholesterol 1 100 cm of small intestine
Site of primary and secondary bile acid absorption Ileum
Only 50% is returned to liver via enterohepatic circulation. False. 98% to 99%
True or False?
Insoluble and not reabsorbed to any significant extent Litocholic acid
A large fraction of bile salts escaped absorption is False. Only a small fraction
eliminated in the feces. True or false?
Major pathway for the elimination of cholesterol Elimination through feces
Amount of bile acid lost in the feces is synthesized from True
cholesterol to maintain the pool of bile acids about 3.5
grams cycled through the intestine 6 to 10 times. True or
false?
Step where bile acid synthesis is regulated CYP7A1 step
Principal rate-liming step in the biosynthesis of bile acids CYP7A1 reaction
Enzyme is feedback regulated via the nuclear bile acid Farsenoid X receptor (FXR)
binding receptor
Increased size of the bile acid pool in the enterohepatic FXR is activated
circulation CYP7A1 gene is suppressed

Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

Enhance CYP7A1 activity Cholesterol of endogenous and dietary origin

Regulate CYP7A1 activity Insulin
Glucagon
Glucocorticoids
Thyroid Hormone
CLINICAL ASPECTS
Correlates with the incidence of atherosclerosis and Serum cholesterol
coronary heart disease
Inflammatory disease cgaracterized by the deposition of Atherosclerosis
cholesterol and Cholesteryl ester from the plasma
lipoproteins into the artery wall
Major cause of heart disease
Important factors in promoting atherosclerosis Elevated plasma cholesterol level (greater than 5.2
mmol/L)
Independent risk factor Elevated blood TAG
Disease with elevated levels of VLDL, IDL, chylomicron Diabetes mellitus
remnants, LDL accompanied by premature or more severe Lipid nephrosis
atherosclerosis Hypothyroidism
Other conditions of hyperlipidemia
Making the LDL:HDL cholesterol ratio a good predictive Inverse relationship between HDL concentrations
parameter and coronary heart disease
Diet plays a part in reducing serum cholesterol. True or True
false?
Most beneficial in reducing serum cholesterol Substitution of poly and monounsaturated FA
instead of saturated FA
Increase proportion of w6 polyunsaturated FA Corn and sunflower seed oil
Increase concentration of monounsaturated FA Olive oil
W3 fatty acids found also beneficial Fish oils
Increase proportion of saturated FA Butter at
Beef fat
Palm oil
Have greater effect in increasing TAG in blood Sucrose and fructose
Means of the unsaturated FA to lower blood cholesterol Up regulation of LDL receptors by poly and
levels monounsaturated FA

(leads to increase in catabolic rate of LDL)

Has anti-inflammatory and triacylglycerol lowering effects W3 fatty acids
Smaller VLDL particles Saturated FA
Contain more cholesterol
Utilized by tissues in slower rate than even larger particles
Atherogenic
Factors associated with increase plasma FFA and increase Emotional stress
TAGS and cholesterol into circulation in VLDL Coffee drinking

Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023
 CHOLESTEROL SYNTHESIS, TRANSPORT AND EXCRETION (BIOCHEMISTRY) 2019 (REF. HARPERS)

Moderate alcohol consumption can lower incidence of Increase HDL concentrations resulting from
CHD increase synthesis of apo A-I
Changes the activity of Cholesteryl ester transfer
protein
Red wine is beneficial
Effects of regular exercise Lowers plasma LDL but raises HDL
TAG concentrations are reduced due to increase
insulin sensitivity that enhances the expression of
lipoprotein lipase
Lower plasma cholesterol Statins
Prevent heart disease
Inhibit HMG CoA Reductase and up-regulating LDL Ator, simva, fluva, prava
receptor activity
Inhibit absorption of cholesterol by the intestine blocking Ezetimibe
uptake via the Niemann-Pick-C-Like 1 protein
Act mainly to lower plasma TAGS Fibrates
Decrease the secretion of TAGS and cholesterol containing
VLDL by the lier Cloibrate, gemfibrozil, nicotinic acid
Primary disorders of the plasma lipoproteins are inherited. True
True or false?
Causes severe hypercholesterolemia associated with Familial Hypercholesterolemia (FH)
premature atherosclerosis
Defect in LDL receptor
LDL is not cleared from the blood
BASED FROM THE TABLE
No chylomicrons, VLDL, LDL formed due to defective apo B Hyperlipoproteinemias
Low or near absence of HDL Familial alpha lipoprotein deficiency
Tangier disease
Fish eye disease
Apo A-I deficiencies
Hypertriacylglycerolemia due to deficiency of LPL, Hyperlipoproteinemias (Familial Lipoprotein Lipase
abnormal LDP or apo C-II deficiency causing inactive LPL Deficiency type I)
Defective LDL receptor Familial Hypercholesterolemia (type IIa)
Deficiency in remnant clearance by the liver is due to Familial type III hyperlipoproteinemias
abnormality in apo E
Overproduction of VLDL often associated with glucose Familail Hypertriacylglycerolemia type IV
intolerance and hyperinsulinemia
Increased concentrations of HDL Familial Hyperalphalipoproteinemia
Deficiency of the enzyme leads to accumulation of large Hepatic Lipase Deficiency
triacylglycerol-rich HDL and VLDL remnants
Absence of LCAT to block in reverse cholesterol transport Familial LCAT deficiency
Lp(a) consists of 1 mol of LDL attached to 1 mol of apo (a) Familial Lipoprotein (a) excess

Gecale, M.D
Brokenshire College – School of Medicine
Batch 2023