SUBJECT: BIOCHEMISTRY

TOPIC: LIPID METABOLISM 3

(Cholesterol Sources &
Biosynthesis & Degradation)
LECTURER: Dr. Laygo
DATE: November 30, 2010

DIET – (Cholesterol) is found in animal fat

BIOSYNTHESIS
 Primarily synthesized by the LIVER(hepatocytes) 2 Acetyl‐CoA  
from Acetyl CoA (formed from oxidative forms 
decarboxylation of glucose) Acetoacetyl‐CoA 
o Synthesis in: catalyzed by 
Thiolase 
Cytoplasm (cytosol)  Membrane of ER  
Found in the 
 Inhibited by LDL uptake by the LIVER Cytoplasm 
(Cytosol) 
*LDL is formed from the lipoprotein VLDL, carries Addition of 
triacylglycerol(TAG), enters the circulation, TAG is another mole 
acted by an enzyme stimulated by Apo CII(in VLDL) of Acetyl‐CoA 
- activates lipoprotein lipase, release free fatty acid
which is deposited in adipose tissue, VLDL
becomes IDL further degradation Important 
LDL(contains cholesteryl ester) reuptake by intermediate, as 
LIVER receptor mediated endocytosis by Apo B- a precursor of the 
100(ligand) synthesis of 
(In absence of receptors or apo B-100, LDL different 
intermediates 
remains in circulation = hypercholesterolemia) —
lipid metab II     
↓Decrease 
Synthesis of 
DEGRADATION      Cholesterol 
Embedded in  
 Occurs in the LIVER ER membrane 
 Cholesterol (Rate Limiting 
 Not utilized by the cell Step)
 Precursor for steroid hormones synthesis
(Anabolic)
o e.g. glucocorticoids, meneralocorticoids,
sex hormones (androgen/estrogen), Vit. D
(Calcitriol or 1,25-dihydroxycholecalciferol)
 Converted  Bile Acids (Catabolic)
STEP 2 – Formation of Isoprenoid units
(See Figure 26-2, Harper’s Illustrated Biochemistry, p226)
BIOSYNTHESIS of CHOLESTEROL (All Carbon atoms from
Acetyl CoA)
STEP 1 – Biosynthesis of Mevalonate (at next column)
(See Figure 26, Harper’s Illustrated Biochemistry, p225)  

3 phosphorylation reduction  
 Isoprene unit contains (5 Carbons)  steps by utilizing (3)ATPs 
 Terpene 
Polymerization of Isoprene units 
 
 
Isopentenyl  Removal of 1 CO2
Monoterpene  1 isoprene unit**  (decarboxylation) 
pyrophosphate 
Farnesyl  6‐carbon to 5‐carbon 
Sesquiterpene  3 isoprene units 
pyrophosphate 
Triterpene  6 isoprene units  Squalene  Isoprene unit  
**Note: Many references state that Monoterpene is formed  (5 Carbon atoms) 
Pyrophosphate 
with 2 isoprene units NOT 1 isoprene unit (e.i. Geranyl  Isopentenyl pyrophosphate or Diphosphate 
pyrophosphate) contradicting what was indicated in the PPT:  
A Monoterpene  Isopentenyl pyrophosphate   Monoterpene**
(terpenes = contain isoprene units)
BIOCHEMISTRY | LIPID METABOLISM 3: Cholesterol Sources & Biosynthesis & Degradation| 1  
 
STEP 3 – Six Isoprenoid Units Form Squalene In the process, an epoxide intermediate (Squalene epoxide)
(See Figure 26-2: Harper’s Illustrated Biochemistry, p226)
is formed – catalyzed by Squalene monooxygenase/
Squalene epoxidase

 3, 3 – dimethylallyl diphosphate an Isomer joins Lanosterol ‐ 1st intermediate in cholesterol 

Isopentenyl pyrophosphate in a head to tail biosynthesis which contain sterol ring. 
manner/head to tail condensation. 4 rings: 
Hypothetical reactions (on formation of Geranyl pyrophosphate) (3) 6‐membered &  
(1) 5‐membered 
1. Ionization  Condensation
2. Ionization  Condensation  Ionization
*sterol ring = cyclopentanoperhydrophenanthrene ring
 Both molecules (with 5 carbon atoms) form a
/sU-klb-pen-t^-nb-per-hUcdrb-fenc^-thrTn/
Geranyl pyrophosphate (a 10-carbon moiety)
Isopentenyl 
Antifungal – utilized in inhibition of the formation of the
pyrophosphate  epoxide.
STEP 5 – Formation of Cholesterol (19 different steps)
Important characteristics:
 Reduction reaction (utilize NADPH+H+ or NADH+H+)
 Molecular oxygen is utilized
 Cholesterol (27 Carbons)
 Another mole of Isopentenyl pyrophosphate in
head to tail manner will condense with Geranyl o 3 Carbons need to be removed from (30-C
pyrophosphate forming Farnesyl diphosphate with Moiety)
(15 Carbons atoms) 3 isoprene units =  Formate (HCOOH) – step 4
Sesquiterpene.
 Carbon dioxide (CO2) – step 9
 Carboxylic group – step 14

 Important Features of Cholesterol

Carbon Feature
3 Hydroxyl groups
Between 5&6 Double bond
17 Aliphatic side chain
18 and 19 Methyl groups
 2 moles of Farnesyl pyrophosphate (15-Carbon) will
condense in a head to head manner forming Squalene
(30-Carbon moiety ) 6 isoprene units =Triterpene)
STEP 4 – Formation of Lanosterol
(See Figure 26-3: Harper’s Illustrated Biochemistry, p227)
Delocalization of the electrons of Squalene to assume a
shape where electrons can easily move with the use of the
enzyme 2,3- oxidosqualene-lanosterol cyclase.

6 isoprene units 

2,3‐ oxidosqualene‐
lanosterol cyclase 

BIOCHEMISTRY | LIPID METABOLISM 3: Cholesterol Sources & Biosynthesis & Degradation| 2  
 
Hydroxyl group at C3 – Esterification of Fatty Acid Palmitic Utility of NADPH + H+ as source of electrons for reduction
acid (most common fatty acid synthesized by eukaryotes) reaction at:
with the use of enzyme ACAT.
 Steps: 7, 8, 9

BILE ACIDS FROM CHOLESTEROL

 formed from cholesterol in the liver

 stored in the gall bladder in bile as bile salts (sodium

and potassium)

o Precursor molecules for synthesis of steroid

hormones
o Geranyl pyrophosphate, Farnesyl
pyrophosphate utilized in synthesis of:
 Heme A
 Dolichophosphate
 Ubiquinone – (ETC) Coenzyme Q
(channel electrons between complex I
and III and between complex II and III)
 G protein – connected to inner leaflet
of plasma membrane; connection to
ACAT inhibitors act  isoprene units making up prenylated
here. Inhibit  protein (serves as anchor).
formation of   utilized during digestion of fats and other lipid
Cholesterol/  substances (act as detergents)
Cholesteryl esters 
o Emulsification: Lipid droplets  Micelle
Demethylation reaction – liberating the carbon moiety (Carboxylic group &
CO2); can be affected by the anti-fungal.

↓ (See Figure 26-3: Harper’s Illustrated Biochemistry, p227)

 Rate limiting step for bile acid synthesis:

Cholesterol 7-α hydroxycholesterol

7‐α‐hydroxylase 
Attachment of hydroxyl group at Carbon number 7

Lanosterol 19 steps Cholesterol

Release of carbon atoms via:
 Step 4: Formate (HCOOH)
 Step 9: Carbon dioxide (CO2) (at reaction 9 to 10)
 Step 14: Carboxylic group (–COOH)

Further reduction using NADPH+H+ and CoA‐SH 
forms Primary bile acids  
(Cholyl‐CoA & Chenodeoxycholyl‐CoA) 
* These 2 are the ONLY Primary bile acids. 

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↓(See FIGURE 26-7: Harper’s Illustrated Biochemistry, p.231)  Taurodeoxycholate –8%
 Various lithocholate –4%

• Fat digestion products are absorbed in the first 100 cm

of small intestine
o enzymes = hydrophilic
o dietary lipase = hydrophobic
o Emulsification: droplet formation = ↑ Surface Area
o Bile salts and acids = ↓ Lipid Aqueous interface
• The primary and secondary bile acids are reabsorbed
almost exclusively in the ileum returning to the liver by
way of the portal circulation (98 to 99%)
o this is known as the enterohepatic circulation
• Less than 500 mg a day escapes reabsorption and is
excreted in the feces.

↓(See FIGURE 26-6: Harper’s Illustrated Biochemistry, p.230)

Secondary bile acids

 Glycocholic acid
1. condensation reaction from Cholyl CoA
2. Uses: glycine
 Taurocholic acid
1. Taurine is formed by the decarboxylation of 
cysteic acid, which in turn is made by
oxidation of cysteine

oxidation decarboxylation

Bile salts
 Lithocholic acid
o Formed from  detergent character of bile salts is due to the
deconjugation + 7α-dehydroxylation of hydrophobic-hydrophilic nature of the molecules
 Taurocholilc acid,  the presence of hydroxyl (or sulfate) and the terminal
 Glycocholic acid,and carboxyl group on the tail give the molecule its
 Chenodeoxycholic acid. hydrophilic face
 Deoxycholic acid  the steroid ring with its puckered plane provides the
o deconjugation + 7α-dehydroxylation from hydrophobic face
glycocholic acid
Function of bile salts
* deconjugation+7α-dehydroxylation(Catalyzed by microbial enzymes) •emulsification of fats due to detergent activity
•aid in the absorption of fat-soluble vitamins (especially
Bile acids
vitamin K)
•cholic acid is the bile acid found in the largest amount in
bile  Vitamin K – synthesized by normal bacterial flora
•bile acids are converted to either glycine or taurine  (Lipid-soluble Vitamins (A, D, E & K)
conjugates (in humans the ratio of glycine to taurine
conjugates is 3:1) •accelerate the action of pancreatic lipase
•have choleretic action –stimulate the liver to secrete bile
APPROXIMATE COMPOSITION OF BILE SALTS •stimulate intestinal motility
 Glycocholate –24% •keep cholesterol in solution (as micelles)
 Glycochenodeoxycholate –24%
 Taurocholate –12%
 Taurochenodeoxycholate –12%
 Glycodeoxycholate-16%

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  ↑ LDL containing cholesterol ester , coalesce
with lysosome  contents LDL in form of
endosome will be hydrolyzed cholesterol
released in the cell = down regulate synthesis of
protein receptors
Type II-B: ↑ VLDL + LDL; often ↓ HDL; ↑ production of VLDL
+ impaired LDL catabolism (from VLDL)
Type III: ↑ IDL (dysbetalipoproteinemia); abnormal
Allows enzyme to attach and begin digestion. apolipoprotein E; impaired catabolism of IDL; ↑
Interior TAG with bile salts around (facing aqueous cholesterol and triglycerides (formerly known as
medium) allowing the enzymes to attach & start digestion broad beta disease)
process. Type IV: ↑ VLDL; often ↓ HDL; impaired VLDL catabolism;
GALLSTONE THERAPEUTIC AGENTS dietary indiscretion ( formerly known as
hyperprebetalipoproteinemia)
•chenodeoxycholic acid (chenodiol; Chenix)
Type V: ↑chylomicrons + VLDL; ↓HDL; ↓lipoprotein lipase +
•ursodeoxycholic acid (ursodiol; Actigall) VLDL hypersecretion (formerly known as mixed
•MOA (Mechanism of Action): lipemia)
–↓ hepatic secretion of cholesterol into bile Factors promoting elevated blood lipids
–inhibition of HMG-CoA reductase (most important enzyme •age
in cholesterol biosynthesis) = inhibit cholesterol –men >45 years of age; women > 55 years of age
biosynthesis
•family history of CAD (Coronary Artery Disease)
–↑ cholesterol solubility
•smoking (α1-antitrypsin inhibits elastase  elastin)
Chenodiol and Ursodiol
Oxidizes α1-antitrypsin specifically its methionin
• both are effective in dissolving cholesterol stones in residue  methionin sulfoxide = α1-antitrypsin
some patients release from normal inhibition of elastase
• ursodiol is the 7-beta epimer of chenodiol •hypertension >140/90 mm Hg
• most effective in dissolving small (<5 mm) floating stones •low HDL cholesterol
in a functioning gallbladder
•obesity > 30% overweight
• cannot dissolve stones that are more than 4% calcium by
weight •diabetes mellitus
Atherosclerosis •inactivity/ lack of exercise
• hardening of the arteries due to the deposition of Mechanisms of action of drugs
atheromas •bind to bile acids/cholesterol
o Atheroma: lipid deposits in the intima of arteries –inhibit absorption/reabsorption
• heart disease is the leading cause of death •increase peroxisomal FA oxidation
• caused by the deposition of cholesteryl esters on the •stimulate lipoprotein lipase
walls of arteries
•inhibit triglyceride lipase
• atherosclerosis is correlated with ↑ LDL (bad
cholesterol*) and ↓ HDL (good cholesterol) •inhibit HMG-CoA reductase
* oxidized form of LDL= harmful •stimulates microsomal 7-α hydroxylase
Drug Classification
•systemic/non-sytemic
•cholesterol lowering agents
–bile acid sequestrants
–sitosterols*
–probucol*
–d-thyroxin*
–HMG-CoA reductase inhibitors

Frederickson – WHO classification •mixed activity (nicotinic acid)

Type I: ↑ chylomicrons, ↓ HDL, absence of lipoprotein lipase; •triglyceride lowering agent

deficiency of apo CII activates lipoprotein lipase. –clofibrate (Atromid-S)
A deficiency of Apo CII will result in accumulation
–gemfibrosil (Lopid)
of chylomicrons and triacylglycerols
(Hyperchylomicronemia) –fenofibrate (Tricor)
Type II-A: ↑ LDL; ↓catabolism of LDL * No longer available commercially in the U.S
(Receptor deficiency or polygenic)
 Receptors synthesized thru Transcription &
Translation
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 BILE SEQUESTERING RESINS 3. Gene expression: cholesterol levels control the
amount of mRNA
 Cholestyramine
 Down regulation process of transcription and
 Colesevelam (WelChol) translation (↓synthesis of receptors and enzyme
molecules)
o polyalkylamine hydrochloride) cross linked with 
epichlorohydrin and alkylated with 1‐bromodecane 
and (6‐bromohexyl) trimethylammonium bromide 
o available as a 625 mg tablet 
o same mechanism of action as colestipol and 
cholestyramine 

 Colestipol (Colestid)
They try to bind bile acids to intestine preventing
absorption into enterohepatic circulation  more
cholesterol transform to bile acids.
 Po (per orem/per os - via the mouth; orally), safest, non
systemic Left side
 bind to bile acids and inhibit reabsorption reactions: Right side
reactions:
 ↑ 7-α hydroxylase activity leading to cholesterol Glucagon &
degradation Epinephrine Insulin
 ↓plasma LDL
 problems:
– abdominal discomfort, bloating, constipation
– decreases drug absorption; wait 4 hrs after
administration of BAS to give drugs
 drug interactions (decreased serum level)
 aspirin
 clindamycin
 clofibrate
 furosemide Left side reactions
 glipzide  During starvation (low energy level)
 tolbutamide
 phenytoin  Predominance of Glucagon and Epinephrine
 imipramine  Inhibits Cholesterol biosynthesis
 methyldopa
 HMG-CoA reductase inactivates when it is
 nicotinic acid
phosphorylated with the use of an ATP and a kinase
 penicillin G HMG-CoA reductase kinase which also requires an
 propranolol activation by coenzyme HMG-CoA reductase kinase
 tetracycline kinase and phosphate from ATP.
 thiazide diuretics
 digoxin  HMG-CoA reductase kinase kinase attaches a
phosphate from ATP to  HMG-CoA reductase kinase
 hydrocortisone
(becomes activated) attaches another phosphate from
 phosphate supplements
ATP to  HMG-CoA reductase (becomes inactive) =
Inhibition of cholesterol biosynthesis
HMG CoA reductase
Right side reactions
 3 different regulatory mechanisms are involved:
1. Covalent modification: phosphorylation(attachment  Insulin predominance
of phosphate, predominating glucagon and
 Allows cholesterol biosynthesis
epinephrin) by cAMP-dependent protein kinases
inactivate the reductase. This inactivation can be  (right top of the figure) HMG-CoA reductase kinase
reversed by 2 specific phosphatases phosphatase by dephosphorylation inactivates
HMG-CoA reductase kinase by removing the
Lipid is catabolized and not synthesized during
phosphate with the use of H2O
starvation/in need of energy.
 (at the bottom of the figure) HMG-CoA reductase
Predominance of Insulin: remove phosphate group phosphatase by dephosphorylation activates HMG-
 activity is stimulated  synthesis of cholesterol CoA reductase by removing also the phosphate with
2. Degradation of the enzyme–half-life of 3 hours and the use of H2O = allowing biosynthesis of cholesterol.
the half-life depends on cholesterol levels STATINS
↑ LDL endocytosed (receptor mediated  Mevastatin
endocytosis) coalesce with lysosome  degrade
contents, release free cholesterol  triggers enzyme  Lovastatin (Mevacor)
degradation  Simvastatin
Protein molecules degraded thru  Pravastatin
Ubiquitination/Ubiquitin cycle

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 Synthetic Statins: o inhibits TG synthesis
 Fluvastatin o stimulates catabolism of VLDL
 Cerivastatin  Indicated primarily for hypertriglyceridemia
Statin group of drugs – inhibitors for HMG-CoA reductase  Same side effects and precaution as in other fibric
acid compounds
 These drugs are administered during mal-elevation of
serum amino transferases: alanine  Half-life: 20 hours
aminotranferease (ALT) and aspartate  Dose: 67-201 mg/day with meals
aminotransferase (AST)
*AST also = SGOT (serum glutamic-oxaloacetic transaminase)
 Normally found in liver, heart, muscle, kidney, and brain Nicotinic Acid (Niacin)
*ALT also = SGPT (serum glutamic-pyruvic transaminase)
 A water soluble vitamin of the B family; nicotinamide is
 Normally found largely in the liver
Increased Serum amino transferases released in the blood are not active (vitamin B3)
indicators of liver damage.  Once converted to the amide, it is incorporated into
NAD (NADH/NADPH)
Precaution:
•mild elevation of serum aminotransferase (should be  In order to be effective, it has to be dosed at the rate of
measured at 2 to 4 month intervals) 1.5 to 3.5 gm daily.
 A sustained release dosage form is available
•minor increases in creatine kinase (myopathy, muscle
pain and tenderness)  Adverse effects:
o GI disturbances (erosion and ulceration)
•do not give during pregnancy
o red flush especially in the face and neck area
o caused by vasodilation of capillaries

FIBRIC ACID DERIVATIVES MOA

 dual plasma triglyceride and cholesterol lowering
Gemfibrosil (Lopid) o decreases VLDL and LDL
MOA  decreases TG lipase in adipose tissue
 stimulates lipoprotein lipase  increases lipoprotein lipase in adipose tissue
 interact with PPAR(peroxisome proliferator-activated Precaution
receptors)  transient cutaneous flush
 inhibits triglyceride lipolysis in adipose tissue
 histamine release
 decreases FFA uptake by the liver
 potentiates BP effect of antihypertensives
 decreases hepatic VLDL/TG synthesis
 slight cholesterol lowering effect Rosuvastatin (Crestor)
Precautions New statins: rosuvastatin (ZD4522)
 similar to clofibrate  Nicknamed” superstastin/ gorilla statin” because of
 myositis (voluntary muscle inflammation) its powerful effect on LDL cholesterol
 GI (indigestion, abdominal pain, diarrhea) Ezetimibe (Zetia)
 cholelithiasis (increased cholesterol biliary secretion)
Half life: 1.1 hours This drug blocks the intestinal absorption of cholesterol. A
dose of 10 mg qd leads to a 19% reduction of LDL; shows
real promise in combo product with statins (Schering-
Clofibrate (Atromid-S)
Plough and Merck)
Primary activity on triglycerides
INVESTIGATIONAL DRUGS
MOA:
 ↑ lipoprotein lipase Acyl-CoA Cholesterol Acyltransferase inhibitors (ACAT
 ↓ VLDL Inhibitors)
 ↑ peroxisomal FFA oxidation –Orphan nuclear receptors:
 inhibits cholesterol biosynthesis
 ↑ biliary secretion of cholesterol •LXR –“oxycholesterol receptor” ---enhanced
cholesterol efflux
Ancillary (secondary effect):
 decreases platelet adhesiveness/fibrinogenbad effect •FXR –“bile acid receptor” ----decreased cholesterol
conversion to bile salts
Precautions
 enhances coumarin activity •Avasemibe (CI-1011)
 renal/hepatic injury contraindication *These drugs do not have a common name yet.
 pregnancy/nursing
 cholelithiasis Squalene synthase inhibitors
 most commonly reported ADR are GI related  squalestin 1, a fermentation product derived from
 liver malignancies (not very common; but has led to Phloma species (Coelomycetes)
scant usage)
 a potent inhibitor of squalene synthase
Fenofibrate (Trecor)
 produces a marked decrease in serum cholesterol and
 A relatively new fibric acid derivative (micronized apoB levels
form of the drug)
 may represent an alternative clinical therapy to
 Lowers plasma TG hypercholesterolemia ║END OF TRANSCRIPTION.
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