P.B. TELEFO, Ph.D.

Senior Lecturer of Biochemistry

UNIVERSITY OF DSCHANG, Cameroon
 Fatty Acids (FA)
LIPIDS PROTEINS LIPOPROTEINS

Cholesterol (CHOL)
MAIN LIPIDS IN
PLASMA TGs
Triglycerides (TGs)
La
yer CHOL-esters
of
PL
Pr , C
Phospholipidsote (PL)
H
i n OL
an
d

General structure of a lipoprotein

 Classification of lipids
• Fatty acids (palmitic, linoleic, etc.)
• Glycerol esters (triglycerides)
• Sterols (cholesterol, hormones,
vitamin D)
• Terpenes (vitamins A, E, K)
• Sphingosine derivatives
(sphingomyelin)
Alteration in Synthesis High levels of TGs and
or Degradation of especially CHOL in the
lipoproteins blood

Cardiovascular
Diseases Atherosclerosis

Cerebrovascular Heart attack

diseases
 Cardiovascular Biggest cause of death
diseases worldwide

WHO

17 million death ∕year 25 million death in

(30% world mortality rate) 2020
Clinical importance of fatty acids

• Fecal fatty acids are sometimes

measured to detect malabsorptive
and pancreatic disorders—the test
is mostly considered obsolete
• Serum free fatty acids help
distinguish between
hyperinsulinemic hypoglycemia
(FFA normal) and disorders of fatty
acid oxidation (FFA elevated and
negative ketones)
Glycerol esters (acylglycerols)
H H

H C OH H C O CO (CH2)n CH3

H C OH H C O CO (CH2)n CH3

H C OH H C O CO (CH2)n CH3

H H

Glycerol Triglyceride

• Triglycerides are the most abundant

family of lipids in plant and animal
cells, and are major components of the
the human diet
 Sterols (cholesterol)

12 17
11 13
C 14 D 16
15
1 9
2 10 8
A B
3 5 7
4 6
HO

• Sterols are steroid backbones that have a

hydroxyl group at position 3 and a
branched aliphatic chain of 8 or more
carbons at position 17
 Cholesterol biosynthesis
• About 2% (approximately 1 g) of total body
cholesterol is replenished each day
– Dietary sources account for less than half
– Cholesterol is synthesized from Acetyl CoA
– 90% of in vivo synthesis occurs in the
intestine and liver (although all cells have the
capability)
• Absorption of dietary cholesterol appears
to have a maximum of approximately 1
g/day
 Cholesterol biosynthesis

Acetyl-CoA 3-Hydroxy-3-methylglutaryl-CoA
HMG-CoA reductase

Squalene Mevalonate

Cholesterol + Lecithin Cholesterol ester

LCAT

“Statin” drugs inhibit this enzyme

 Measuring triglycerides
(reference method)

Triglycerides
fatty acids + glycerol
KOH
formic acid + formaldehyde
Periodate

chromogen
chromotropic acid
=570 nm
• Triglycerides are extracted into
chloroform prior to analysis
 Measuring triglycerides
(enzymatic method)
Triglycerides
Lipase
Glycerol + FFAs
Glycerokinase
ATP Glycerophosphate + ADP
Glycerophasphate
oxidase
Dihydroxyacetone + H2O2
Quinoneimine dye
max 500 nm Peroxidase
 Measuring cholesterol by L-B

L-B reagent
H2SO4/HOAc
HO HOO2S

Cholesterol Cholestahexaene sulfonic acid

max = 620 nm

• The Liebermann-Burchard method is used by the

CDC to establish reference materials
• Cholesterol esters are hydrolyzed and extracted into
hexane prior to the L-B reaction
 Enzymatic cholesterol methods
Cholesterol esters
Cholesteryl
ester
hydroxylase Cholesterol
Cholesterol
oxidase
Choles-4-en-3-one + H2O2
Phenol
4-aminoantipyrine
Quinoneimine dye Peroxidase
(max500 nm)
• Enzymatic methods are most commonly adapted to
automated chemistry analyzers
• The reaction is not entirely specific for cholesterol, but
interferences in serum are minimal
 Liver, Skeletal
Intestine muscle and
Adipose tissues

Liver body’s Cells

Adipose
Liver
tissues

Body’s
tissues Liver
 Lipoproteins
• In order to be transported in blood, lipids must
combine with water-soluble compounds, such
as phospholipids and proteins.
Density (g/ml) Class EM Diameter (nm) Composition Apolipoproteins Origin
100-1000 90% TG 2% Intestine
Apoproteins
<0.95 CM No migration
5% Apo AI AII
phospholipids
3% cholesterol AIV B48 (C and
esters E)
<2% protein
VLDL 30-80 60% TG 5-10% Liver
Apoproteins
0.95-1.006 Pre-beta
15% B 100
20% cholesterol C (E)
10% protein
IDL Broad-beta 25-50 50% TG E,B Catabolism of
50 cholesterol VLDL
1.006-1.019
18% protein
18-28 10% TG 22% Catabolism of
LDL Apoproteins VLDL via IDL
1.019-1.063 beta B100
20-25%
phospholipids
70% CHOL esters
25% protein

5-15 5% TG 45-50% Liver, intestine,

HDL Apoproteins catabolism of
>1.063 alpha
25%
A1 , AII CM and VLDL
phospholipids
20% CHOL esters
33% protein C (D, E)
 BY DENSITY

From larger and less dense to smaller and denser

 Lipoprotein classes
%TG %Chol LPE
Chylomicrons 86 3 Origin
VLDL 55 12 Pre-
IDL 23 29 Pre-/
LDL 6 42 
HDL 3 15 
Lp(a) (LDL) (LDL) Pre-
 Appearance of hyperlipidemia
• Standing Plasma Test for chylomicrons
– Plasma is placed in refrigerator (4°C)
overnight
– Chylomicrons accumulate as floating
“cream” layer
– Chylomicrons in fasting plasma are
abnormal
Lipoprotein electrophoresis
 Pre- 

- Migration +

Chylomicrons LDL IDL VLDL HDL

Lp(a)

• LEP is no longer a common laboratory

test
– Standing plasma test for chylomicrons
– Total cholesterol, TG, HDL, and LDL can
be measured directly
 Fredrickson classification
Type Refrig. LPE LPs
I Pos, clear Normal TG (chylos)
IIa Neg, clear   band LDL
IIb Neg, cloudy  , pre-  LDL, VLDL

III Occ.,  pre-  Chol, TG, VLDL

cloudy
IV Neg, cloudy -2 VLDL

V Pos, cloudy -2 VLDL Chylos

 Measuring HDL cholesterol
• Ultracentrifugation is the most accurate
method
– HDL has density 1.063 – 1.21 g/mL
• Routine methods precipitate apolipoprotein B
with a polyanion/divalent cation
– Includes VLDL, IDL, Lp(a), LDL, and chylomicrons

Dextran sulfate
HDL, IDL, LDL, VLDL HDL + (IDL, LDL, VLDL)
Mg ++

• Newer automated methods use a modified form of cholesterol

esterase, which selectively reacts with HDL cholesterol
 Indirect LDL cholesterol
• Friedewald formula assumes that all
cholesterol is VLDL, LDL, and HDL
lipoproteins
– Chylomicrons are usually low in normal,
fasting subjects, and IDL and Lp(a) are
usually insignificant contributors to total
cholesterol
• Since VLDL is 55% TG and 12% Chol:
[LDL Chol] = [Tot Chol] – [HDL Chol] –
[TG]/5
 Direct LDL cholesterol
• Older direct methods for LDL involved
precipitation with heparin or polyvinyl
sulfate
• Newer methods involve precipitation of
VLDL, IDL, and HDL with polyvalent
antibodies to Apo A and Apo E
– LDL is almost exclusively Apo B-100
 Direct vs. Indirect LDL
• The Friedewald equation assumes that
chylomicrons, IDL, and Lp(a) are not
significant
– Non-fasting specimens can have
chylomicrons
– TG > 400 mg/dL indicates the presence of
chylomicrons (or remnants)
• Type III hyperlipidemia is
characterized by high -VLDL, which
has a 3:1 TG:C ratio
 Apolipoproteins

• The protein composition differs from

one lipoprotein class to another, and
the protein constituents are called
Apolipoproteins
Functions of apolipoproteins

• Activate enzymes involved in lipid

metabolism (LCAT, LPL)
• Maintain structural integrity of
lipid/protein complex
• Delivery of lipids to cells via
recognition of cell surface receptors
Apolipoprotein content of LPs
Lipoprotein Apolipoprotein(s)
Chylomicron AI, B-48, CI, CII, CIII

VLDL B-100, CI, CII, CIII, E

IDL B-100, E
LDL B-100
HDL AI, AII
Lp(a) (a), B-100
Cholesterol metabolism (exogeneous)
Dietary cholesterol, Apo-C, E from HDL
triglycerides

Endothelium
A A

LPL
C,TG C,TG C
E
B B
Chylomicron

C,TG
E
B
Hepatocyte B/E receptors Chylomicron
remnant
 Cholesterol metabolism
(endogeneous)

Endothelium
LPL
C,TG C
E
B
VLDL
Hepatocyte B-100
receptors

C C,TG
E
B B
LDL IDL
 Nascent AI
discoidal ApoAI R
HDL E LIVE
ApoE
SR-B1
Chylomicrons
+ LCAT FC
VLDL
Ap
oC
Ap
oA
I

AI

LCAT LCAT
E
C

HDL3 Hepatic Lipase Triglyceride

Spherical
HDL2
 HDL
B100 B100
TAG TAG
+ +
CHOL CHOL CII
E
CII E
Nascent VLDL Mature VLDL

L
LP
LRP LPL
B100
TAG
LIVER +
CHOL CII
LDLR E

IDL Glycerol
HL HL +
Fatty acids
B100
CHOL
CII
E Peripheral
LDL LDLR tissue
Atherosclerosis (ASVD):
TheWhen LDLsLDLs
modified accumulate, they become
also promote oxidized by free
the differentiation radicals.
of monocytes
 Artery wall thickens as a result of the accumulation of cholesterol.
Themacrophages.
into modified LDLsMacrophages,
then stimulate in endothelial cells to express
turn, express scavengera
 A syndrome affecting arterial blood vessels, a chronic inflammatory
protein, to
receptors Monocyte Chemotactic
take up the modified Protein-1 (MCP-1),
LDLs, resulting thatformation
in the attracts
response, caused by the accumulation of macrophages and promoted by
of monocytes
lipid-filled from
foam the blood
cells, into the artery
the hallmark wall.
cells of atherosclerosis
LDL without adequate removal of CHOL by HDL.
 NORMAL ATHEROSCLEROTIC
A ARTERY
B
ARTERY

PLA
QUE

NARROWED
ARTERY

NORMAL CROSS SECTION

EPIDEMIOLOGY

High concentrations of HDL (over 60 mg/dL) is a protective value

Accelerated
Atherosclerosis
deposition
typicallyofbegins
cholesterol
in early
in the
adolescence,
walls of arteries
and is
against cardiovascular diseases. Low concentrations of HDL (below
found
leads to
in atherosclerosis,
major arteries, the
yet underlying
is asymptomatic
causeand
of cardiovascular
not detected
40 mg/dL for men, below 50 mg/dL for women) increase the risk for
by
disease.
most diagnostic
atherosclerotic diseasesmethods during
(Barter et life.
al., 2007).

AG
E
Increased
Atherosclerosis
Risk
 Familial Hypercholesterolemia (FH)

It is a genetic Anatomic
disorder characterized
detection methods
by high
: lipoprotein
cholesterol
subclass
levels,
specifically very analysis,
high levels
abdominal
of LDL, girth
"bad. cholesterol”, in the blood
DIAGNOSIS
and early cardiovascular disease.
Physiologic measurement methods : blood pressure,
elevated blood glucose methods
FH is often associated with the mutation of R3500Q, which causes
replacement of Arginine by Glutamine at position 3500.
Non-pharmaceutical means: cessation of smoking ,
practicing regular exercise.
REMEDY AND
The mutation is located on a part of the protein that normally binds with
PREVENTION
Medicines:
the LDL receptor, and binding isstatins...
reduced ashave antioxidant
a result effects and
of the mutation.
have therapeutic success in reducing cardiac
'events'.
 Dyslipoproteinemias
• Causes can be primary or secondary
– Secondary causes include starvation, liver disease, renal
failure, diabetes, hypothyroidism, lipodystrophies, drugs
• Primary causes of hyperlipidemia:
– Increased production
– Defective processing
– Defective cellular uptake
– Inadequate removal
 Dyslipoproteinemias
• Hyperchylomicronemia
– LPL deficiency
– Apo C-II deficiency
 Hyperchylomicronemia
Dietary cholesterol, Apo-C, E from HDL
triglycerides

Endothelium
A A

LPL
C,TG C,TG C
E
B B
Chylomicron

Chylomicrons
Triglycerides C,TG
HDL E
LDL B
Hepatocyte B/E receptors Chylomicron
remnant
 Dyslipoproteinemias
• Hyperchylomicronemia
– LPL deficiency
– Apo C-II deficiency
• Hyperbetalipoproteinemia
– Overproduction of VLDL
– Enhanced conversion of VLDL to LDL
– LDL enriched with cholesteryl esters
– Defective LDL structure
– Decreased LDL receptors
 Hyperbetalipoproteinemia

Endothelium
LPL
C,TG C
E
B
VLDL
Hepatocyte B-100
receptors

LDL C C,TG
E
Normal TG B B
LDL IDL
 Dyslipoproteinemias
• Combined hyperlipoproteinemia
– Normal LDL receptors
– Overproduction of VLDL and Apo B-100
 Combined hyperlipoproteinemia

Endothelium
LPL
C,TG C
E
B
VLDL
Hepatocyte B-100
receptors

LDL C C,TG
E
Normal TG B B
LDL IDL
 Dyslipoproteinemias
• Combined hyperlipoproteinemia
– Normal LDL receptors
– Overproduction of VLDL and Apo B-100
• Dysbetalipoproteinemia
– Both cholesterol and triglyceride elevated
– Mutant form of Apo E
 Dysbetalipoproteinemia
Dietary cholesterol, Apo-C, E from HDL
triglycerides

Endothelium
A A

LPL
C,TG C,TG C
E
B B
Chylomicron

Cholesterol
TG C,TG
E
B
Hepatocyte B/E receptors Chylomicron
remnant
 Dyslipoproteinemias
• Familial hypercholesterolemia
– Defect in LDL receptor
• Absent
• Defective
– Incidence = 1:500
 Familial hypercholesterolemia

Endothelium
LPL
C,TG C
E
B
VLDL
Hepatocyte B-100
receptors

LDL C C,TG
E
or n TG B B
HDL LDL IDL
 Dyslipoproteinemias
• Familial hypercholesterolemia
– Defect in LDL receptor
• Absent
• Defective
– Incidence = 1:500
• Familial defective Apolipoprotein B-100
 Familial hypercholesterolemia

Endothelium
LPL
C,TG C
E
B
VLDL
Hepatocyte B-100
receptors

or n LDL C C,TG

E
B B
LDL IDL
 High cholesterol, high LDL
• Diet/Lifestyle
• 2° to hypothyroidism or nephrotic syndrome
(disruption of Apo-B metabolism)
• Polygenic: (means we don’t know)
• Familial hypercholesterolemia
• Familial defective Apo-B
• Rare disorders
 High TG, normal cholesterol
• Diet/Lifestyle
• 2° to diabetes, thiazide diuretics, Cs, beta-
blockers, CRF/Nephrotic syndrome
• Familial hypertriglyceridemia (etiology
unknown)
• ApoC-III excess (interferes with LPL)
• LPL deficiency
• ApoC-II deficiency
 High cholesterol, TG
• Obesity
• 2° to steroids, Cs, hypothyroidism, CRF
• Familial combined hyperlipidemia
(multifactorial)
• Peroxisome proliferator-activator receptor
• Dysbetalipoproteinemia (Type III)
• Hepatic lipase deficiency (rare)
 Low cholesterol, low/normal HDL
• Abetalipoproteinemia (ApoB degraded after
synthesis causes fat malabsorption)
• Hypobetalipoproteinemia (genetically
defective ApoB)
• Chylomicron retention disease (unknown
cause)
 Low HDL

• Lifestyle
• 2° to steroids, beta-blockers, progestogens
• Familial hypoalphalipoproteinemia (ApoA-I,
C-III, or A-IV defects)
• ApoA-I variants
• Tangier disease (enhanced HDL degredation)
• LCAT deficiency
 High HDL
• Lifestyle (ethanol)
• 2° to phenytoin, phenobarbitol, rifampicin (p-
450 inducers) and estrogens
• Cholesteryl Ester Transfer Protein defects