Plasma Lipids, Lipoproteins, Apolipoproteins

Lecture Outline
• Plasma lipids – function, biochemistry, groups
• Cholesterol, fatty acids, triglycerides, phospholipids
• Lipoproteins
• Classification of lipoproteins
• Apolipoproteins
• Low density lipoprotein
• Control of LDL/cholesterol
• Factors influencing plasma levels of LDL
• Lipoprotein receptors - LDL receptor
• Exogenous lipid pathway
• Endogenous lipid pathway
• Very low density lipoprotein
• Reverse cholesterol transport
• High density lipoprotein
• Enzymes involved in lipid metabolism
• Looking at the pathways and lipoproteins
 Lipids
• Endogenous: synthesized by the body
• Exogenous: derived from food
• Lipids needs to be transported from one organ to
another
• Lipids are insoluble in water
∴Circulated in body fluids as soluble protein
complexes
 Lipids - Functions
• Metabolic fuel
• Hormones & hormone precursors
– Sex hormones
– Glucorticoids
– Mineralocorticoids
• Aiding in digestion – bile lipids
• Energy storage
• Functional and structural components of cell
membranes
– Forming membrane
– Allow nerve conduction
• Fat-soluble vitamins
 Lipid Biochemistry

• Insoluble in H2O
• Soluble in organic solvent
• Chemically, lipids are either:
– Components that yield fatty acids on hydrolysis,
or
– Complex alcohols that combine with fatty acids to
form ester
• Some contain non-lipid groups such as sialic,
phosphoryl, amino or sulfate groups
– Increase solubility
 Plasma Lipids
• Divided into groups based on chemical
structure:
1. Cholesterol
2. Fatty acids
3. Triglycerides
4. Phospholipids
 Cholesterol
• Steroid precursor to bile acids, steroid hormones
• Cholesterol level in body = 150 – 200 mg/dl
• Sources
– Diet: 400 – 700 mg/day
• On average 30 – 60% absorbed
– De novo synthesis
– Synthesis in intestines and liver
• The liver
→ HDL, VLDL
→ Free cholesterol in bile
→ Conversion → bile salts
• Excretion = 1 g/day as bile acids
 Cholesterol
• Can be esterified by:
– LCAT (lecithin cholesterol acyl-transferase) in
plasma
– ACAT (acyl cholesterol acyl-transferase)
intracellularly
 Cholesteryl ester
– Packed into lipoprotein particles
– Transported
• 1/3 of daily cholesterol production –
catabolised to bile acids
• Bile acids are important in absorption
of cholesterol
 Fatty Acids
• Straight chain carbon compounds
• CH3(CH2)nCOOH
 Fatty Acids
• Esterified with glycerol → triglycerides
• Non-esterified (NEFA) or free (FFA)
• Oxidised for production of energy in mitochondria by β
oxidation
• During prolonged starvation excessive degradation of
fatty acids by β oxidation in liver causes an acetyl CoA
excess
• Acetyl CoA acetoacetic acid
∀ β hydroxybutanic acid acetones are produced
(ketone bodies)
• Excess production of ketone bodies → ketosis
 Plasma Fatty Acids

• In plasma – free fatty acids are carried bound to

albumin
• Free fatty acids: immediate source of energy → yield
large quantities of ATP
 Triglycerides
• Fatty acid esters of glycerol containing 3 different fatty
acids
• Transported as lipoproteins from gut and liver to
tissues
• After hydrolysis → fatty acids taken up, re-esterified
and stored as triglycerides
 Phospholipids

• Complex lipids that resemble triglycerides but contain

phosphate + 1 nitrogen base in place of one fatty acid
• Important components of cell membranes and
lipoproteins – maintain solubility of non-polar lipids
and cholesterol
 Lipoproteins

• Lipids + proteins
• Synthesis: liver and intestines
• Modified after secretion by enzyme-catalysed reactions
• Remnants are taken up by receptors on cell surfaces
• Modification and uptake of remnants – regulated by
apolipoproteins ~ the protein component
• Lipoproteins are classified by density, which in turn
reflects their size
• The more lipid a complex contains, the greater the
lipid/protein ratio → the larger it is, and the lower the
density
 Classification of Lipoproteins
Lipoproteins Role
Chylomicrons Transport exogenous lipids from intestines to all
cells, large, triglyceride rich
VLDL Transport endogenous lipids from liver to cells
(very low density Large, triglyceride rich
lipoproteins)
IDL Transient intermediate formed during
(intermediate density conversion of VLDL → LDL, undetectable in
lipoproteins) normal plasma
Cholesterol + triglycerides
LDL Formed from VLDL
(low density Transport cholesterol to cells
lipoproteins) Contains mostly cholesterol
HDL Transport of cholesterol from cells to liver
(high density
lipoproteins)
 Lipoproteins - chylomicrons

• Dietary fat → fatty acids + cholesterol

• + cholesterol (from bile)
• Re-esterification (in intestinal mucosal cells) →
tryiglycerides + cholesteryl esters
• + phospholipids + apoA + apoB ⇒ chylomicrons
• Chylomicron secretion from cells into lymphatic system
depends on presence of apoB
• + apoC + apoE in lymphatic system and in plasma
Tissue of origin and function of plasma lipoproteins

Origin Functions
Chylomicrons Intestines Absorption of dietary
fat
Chylo. remnants Plasma Deliver dietary fat to
liver
VLDL Liver Deliver triglycerides from
liver to other tissues
IDL Plasma Initial product of
VLDL catabolism
LDL Plasma Cholesterol transport
HDL Liver, Removal of excess
intestines cholesterol from
tissues and lipoprotein
 Apolipoproteins

• Lipid-binding protein constituents of plasma

lipoproteins
• Transport dietary lipids from intestines to liver, and
endogenously synthesized lipids from liver to
storage tissues
• Apolipoproteins control:
– Normal lipid secretion
– Activation of enzymes concerned with lipid
metabolism
– Receptor recognition of lipoproteins
 Apolipoproteins

• Apoliproteins are designated by letters, e.g. apoA

• If each class has more than one member, then
Roman numerals are used:
apoA-I, apoA-II, apoB-48, apoB-100,
apoC-I, apoC-II
 Apolipoproteins

Lipoproteins Apolipoproteins

Chylomicrons B-48, A-1, A-IV, C-II, E

VLDL B-100, E, C-I, C-II

ILDL B-100, E, C-II

LDL B-100,

HDL A-I, A-II, C-II, E

 Apolipoproteins - Functions
Apolipoproteins Function

A-I Cofactor for the enzyme LCAT

(lecithin-cholesterol acyltransferase)
B Secretion of chylomicrons and VLDL
Binding of LDL to receptor
C-II Cofactor for lipoprotein lipase

E Binding of IDL and chylomicron

remnant particles to receptor
 LDL

• Small, cholesterol-rich lipoprotein containing only

apoB
• Longer life than its precursors (VLDL and IDL)
∀ ≈ 70% of total cholesterol in plasma
• Taken up by specific receptors on cell surfaces – LDL
receptors or apoB/E receptors
• LDL receptors present on all cells and most abundant
in liver
• After entering cells → LDL broken down by lysosomes
→→ cholesterol released
 Control of LDL / Cholesterol level

• Most cells can synthesise cholesterol but cholesterol

taken up by receptors inhibit intracellular cholesterol
synthesis and prevents further uptake
• One way to control uptake of LDL by cells is to reduce
synthesis of LDL receptors
• Most plasma LDL is removed by LDL receptors
• If plasma LDL level is high, some LDL may enter cells
by a passive unregulated route – possible because of
its small size
 Factors influencing plasma level of LDL

• Plasma level of LDL and cholesterol determined by the

rate of uptake by LDL receptors
• Liver has the central role:
– Contains the most of the LDL receptor in the body
– Synthesizes most of the endogenous cholesterol
– Receives cholesterol from the diet and from
lipoproteins
– Is the only organ that can excrete cholesterol from
the body
 LDL receptor
• Down regulated when sufficient cholesterol is available
• Up regulated when cells require additional cholesterol
• Regulation is controlled either by:
– Extent to which existing receptors are recycled to the
cell surface after endocytosis, or
– The amount of new receptors that are synthesized
• Concentration of LDL receptors on hepatic cell surface
depends on the amount of cholesterol in cells
• Increase in intracellular cholesterol → reduction in
number of receptors
• Factors that increase cholesterol in liver will increase
plasma LDL concentration because number of receptors
is reduced
 LDL receptor

• Best characterised
• Present in liver and some tissues
• Contained in coated pits
• 160,000 Dalton glycoprotein
• Recognises apoB-100 and apoE
• Through binding of apoB-100 – mediates clearance of
IDL
• VLDL contains both apoB-100 and apoE but cannot
bind because VLDL also contains apo C-III
• Expression is regulated by the need of the cell for
cholesterol
 Lipoprotein receptors
Receptor Recognition LP Tissue

LDL apoB-100 LDL Liver

apoE IDL Other

Chylomicron apoE Chylomicron Liver

remnants remnants

IDL Unknown HDL Liver

Scavenger Chemically Modified Endothelium

modified or damaged macrophages
apoB-100 LDL
 Lipoprotein receptors
Receptor Role

LDL Removal of IDL, LDL from circulation

Mediate delivery of cholesterol from plasma
to tissue
Chylomicron Uptake of dietary fat by the liver
remnants Also called the apoE receptor

IDL HDL binding to cells

Existence is questionable

Scavenger Removal of chemically altered LDL

Poorly defined
 Overview of lipoprotein function

Dietary fat transport Triglyceride secretion Reverse cholesterol

system system transport

Intestine Liver Extrahepatic

IDL tissues
Chylomicrons LDL
HDL
Chylo VLDL LDL
remnants
Extrahepatic
Extrahepatic Liver
tissues Liver
tissues
 Lipoproteins
• When a lipoprotein reaches its destination:
– Chylomicron and VLDL carry triglycerides/cholesterol
– Lipoprotein docks via apoC-II
– Activates lipoprotein lipase
– Releases fatty acids absorbed by cells
– Some will complex to serum albumin for further
transport
– Remainder of VLDL is now IDL and returned to liver,
or IDL is further degraded by lipase to LDL
– LDL returns to liver for endocytosis or endocytized by
peripheral cells (must have LDL receptor)
– Remainder of chylomicron-chylomicron remnant is
returned to liver
– Degraded by lysosomes
 Exogenous Lipid Pathway
• Exogenous lipids – lipids released by digestion of dietary
fat (fatty acids, cholesterol) and cholesterol released by
bile
• Digestion: not much in the stomach
• Lipids broken into FFA and acyl-glycerols
• Transported across the intestinal walls
• Triglycerides are reassembled before entering
bloodstream and associated with proteins before being
transported → complexes called chylomicrons
• Upon reaching their destinations triglycerides are:
– Broken down to provide energy
– Stored in adipose tissues
 Endogenous Lipid Pathway

• Lipids synthesized and released by the liver

• Triglycerides from glycerols and fatty acids from fat
stores or synthesized from glucose
• Hepatic cholesterols are either synthesized locally or
derived from lipoproteins e.g. from chylomicron remnants
• Transported from the liver in VLDL
 VLDL

• Large, triglyceride-rich
• Incorporates apoB, apoC, ApoE – and after secretion,
incorporates more apoC from HDL
• In peripheral tissues → hydrolysis by lipoprotein lipase
→ release triglycerides
• After hydrolysis ⇒ IDL
• IDL contains triglycerides + cholesterol + apoB + apoE
• IDL either:
– Taken up by liver
– Loses remaining tryiglycerides and apoE → LDL
 Reverse cholesterol transport
• Cells produce cholesterol needed for cellular
homeostasis
• The liver: the only organ that is capable of degrading
cholesterol
• Cholesterol must be transported through blood to the
liver for processing, degradation, secretion and excretion
into bile
• Since the liver is the center of cholesterol homeostasis in
the body, cholesterol that moves from peripheral tissues
to the liver is considered to be moving in the reverse
direction
 Reverse cholesterol transport

• HDL: the only particle capable of receiving cholesterol

from peripheral cells
• HDL → responsible for most of the reverse cholesterol
transport
• ApoA-I is the major apolipoprotein in HDL and the
primary acceptor for un-esterified cholesterol from
peripheral cells
• Level of plasma apoA-I is lower in smokers than in non-
smokers
• Exercise training increases plasma level of apoA-I
• Inverse relationship between coronary disease and the
plasma apoA-I level
Reverse Cholesterol Transport
Extrahepatic Tissues
C

HDL3
C+FA
LCAT

CE

HDL2 TG VLDL

CE CE
Liver
C
Bile
C
Faeces
 HDL

• Synthesized de novo in liver and small intestines

• Protein-rich disc shaped particles, containing
phospholipid, free cholesterol, apoA, apoB
• Primary apolipoproteins are apoA-I, apoC-I, apoC-II
and apoE
• One major function of HDL is to act as circulating
stores of apoC-I, apoC-II and apoE
 HDL
• HDL → converted to spherical lipoproteins with
accumulation of cholesterol esters
• This addition of cholesterol esters convert nascent
HDLs to HDL2 and HDL3
• Cholesterol-rich HDLs return to liver where they are
endocytosed
• Hepatic uptake of HDL (reverse cholesterol transport)
may be mediated by SR B-I receptor or through lipid-
lipid interaction
• Macrophages also take up HDL through apoA-I
receptor interaction
 HDL
• Since macrophages take up HDL through apoA-I
receptor interaction - HDL can then acquire cholesterol
and apoE from macrophages
• Enriched HDLs are then secreted from the
macrophages
• The added apoE in these HDL lead to an increase in
their uptake and catabolism by the liver
• HDL also acquire cholesterol by extracting it from cell
surface membranes → causing cholesterol stored
intracellularly as cholesteryl esters to be mobilized to
replace the cholesterol removed by HDL
• As a result, intracellular cholesterol level is lowered
 HDL Metabolism
Intestines Liver

Nascent HDL

Tissues C C Chylomicrons
PL VLDL
Apos
HDL3

C LCAT
CE
CE
VLDL
TG
HDL2
TG/CE
Hepatic lipase

LIVER
 Enzymes involved in lipid metabolism
Lipoprotein Lipase
• Hydrolyse triglycerides contained in circulating
chylomicrons and VLDL → monoglycerides and fatty acids
• Acts on surface of capillary endothelial cells activated by
apoC-II (present in VLDL and chylomicron)
• Present in many tissues –adipose tissue, mammary
glands, heart
• Activity increased by insulin, when insulted is administered
• Enzyme formed within the tissue and then secreted to the
surface of endothelial cells
• Binds firmly to the glycoprotein present on the surface of
endothelial cells
• Acts on triglycerides while firmly bound to endothelial
surface
• Monoglycerides are then transported to the liver where
they are degraded by a hepatic monoglycerate lipase
Enzymes involved in lipid metabolism
Hepatic Lipase
• Similar in properties to lipoprotein lipase
• Present in liver sinusoids
• A lipid hydrolase
• Acts on IDL and HDL2
• Acts on triglycerides and also on phosphoglycerides
present on surface coats
• Probably is responsible for the conversion of IDL to
LDL or for processing of IDL for uptake by the liver
• Also may be responsible for HDL2 processing: either
for uptake by liver or for conversion back to HDL3
Enzymes involved in lipid metabolism
Acid Lipase
• Acts intracellularly after lipoprotein is removed from
circulation by receptor-mediated endocytosis
• Contained in lysosomes
• Member of a family of enzymes called lysosomal
acid hydrolases
• Hydrolyses triglycerides and cholesterol esters
remaining in lipoproteins → releasing fatty acids and
cholesterol for utilisation in cells
Enzymes involved in lipid metabolism
LCAT
lecithin-cholesterol acyltransferase

• Mediates cholesterol esterification in plasma

C + FA CE
LCAT

• Intracellularly – acyl-cholesterol acyl-transferase

(ACAT)
• Re-esterify excess cholesterol for intracellular
storage
• Activity is enhanced by presence of intracellular
cholesterol
Enzymes involved in lipid metabolism
CETP

• Cholesteryl ester transfer protein

• Glycoprotein
• Synthesized mainly in the liver
• In plasma, CEPT is predominantly bound to the HDL
containing apoA-I only particles
• Mediates the exchange of HDL cholesteryl esters with
triglycerides of Apo B containing lipoproteins
• Plays a central role in reverse cholesterol transport