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Cholesterol and Steroid Metabolism

I.
Overview
Cholesterol characteristic steroid alcohol of
animal tissues
- Structural component of all cell
membranes (modulate its fluidity)
- Precursor of bile acids, steroid hormones,
and vitamin D (specialized tissues)
Liver regulate bodys cholesterol
homeostasis
Cholesterol sources:
Dietary cholesterol
Cholesterol synthesized de novo by
extrahepatic tissues and by the liver
itself
Fates of cholesterol:
Eliminated from the liver as
unmodified cholesterol in the bile
Converted to bile salts that are
secreted into the intestinal lumen
Component of plasma lipoproteins sent
to the peripheral tissues
Atherosclerosis lipid deposition leads to
plaque formation causing narrowing of
blood vessels
II.
Structure of Cholesterol
Cholesterol very hydrophobic
- Consists of 4 fused hydrocarbon rings
(steroid nucleus)
- Has an eight-carbon, branched
hydrocarbon chain attached to carbon 17
of the D ring
- Ring A has OH at carbon 3
- Ring B has a double bond between carbon
5 and carbon 6
A. Sterols
- Steroids with 8 to 10 carbon atoms in
the side chain at carbon 17 and OH at
carbon 3
Cholesterol major sterol in animal tissues
Plant sterols (e.g. -sitosterol) poorly
absorbed by humans
- After entering enterocytes, they are
actively transported back into the
intestinal lumen
- Reduce absorption of dietary cholesterol
used in dietary treatment of
hypercholesteremia

Commercially available trans fatty acidfree margarine

B. Cholesteryl esters
- Not bound in membranes
- Normally present in low levels in most
cells
- Must be transported in association with
protein as a component of a lipoprotein
particle or be solubilized by phospholipids
and bile salts in the bile
III.
Synthesis of Cholesterol
- Endergonic
- Driven by hydrolysis of the high-energy
thioester bond of acetyl coenzyme A (CoA)
and the terminal phosphate bond of ATP
- Requires enzymes in both the cytosol and
the membrane of the smooth ER
- Responsive to changes in cholesterol
concentration
Cholesterol synthesized by virtually all
tissues in humans
Make the largest contributions to the
bodys cholesterol pool:
Liver
Intestine
Adrenal cortex
Ovaries
Testes
Placenta
Acetate provides all the carbon atoms in
cholesterol
NADPH provides the reducing equivalents
Imbalance in regulation can lead to elevation
in circulating levels of plasma cholesterol with
the potential for vascular disease
A. Synthesis of 3-hydroxy-3methylglutaryl (HMG) CoA
- First 2 reactions are similar in the ketone
bodies pathway
- Result in the production of HMG CoA

First 2 acetyl CoA


molecules condense
to form acetoacetyl
CoA

Final step ER-associated pathway


- Includes several different enzymatic
reactions

Third molecule of
acetyl CoA is added
producing HMG CoA

Smith-Lemli-Opitz syndrome (SLOS)


- Relatively common autosomal recessive
order of cholesterol biosynthesis
- Caused by partial deficiency in 7dehydrocholesterol-7-reductase
- One of several multisystem, embryonic
malformation syndromes associated with
impaired cholesterol synthesis
7-dehydrocholesterol-7-reductase
enzyme involved in the migration of the
double bond

HMG CoA six-carbon compound


Liver parenchymal cells
contain 2 isozymes of HMG CoA
synthase
- Cytosolic enzyme: participates in
cholesterol synthesis
- Mitochondrial enzyme: functions in the
pathway for ketone body synthesis
B.
-

Synthesis of mevalonate
Reduction of HMG CoA to mevalonate
Catalyzed by HMG CoA reductase
Rate-limiting and key regulated step in
cholesterol synthesis
- Occurs in the cytosol
- Uses 2 molecules of NADPH as reducing
agent; releases CoA
- Irreversible
HMG CoA reductase intrinsic membrane
protein of the ER with the enzymes catalytic
domain projecting into the cytosol
C. Synthesis of cholesterol
IPP precursor of a family of molecules with
diverse functions, the isoprenoids
Cholesterol sterol isoprenoid
Nonsterol isoprenoids
e.g. dolichol and ubiquinone
Prenylation covalent attachment of
farnesyl to proteins
- One mechanism for anchoring proteins to
plasma membranes
Squalene formed from 6 isoprenoid units
- 3 ATP are hydrolyzed per mevalonate
residue converted to IPP
Total: 18 ATP required to make the
polyisoprenoid squalene

Mevalonate is converted to 5pyrophosphomevalonate in 2 steps


each of which transfers a phosphate
group from ATP

Isopentenyl pyrophosphate (IPP) is


formed by the decarboxylation of 5pyrophosphomevalonate. The reaction
requires ATP.

IPP is isomerized to 3,3-dimethylallyl


pyrophosphate (DPP)

D. Regulation of cholesterol synthesis


HMG CoA reductase rate-limiting enzyme
- Major control point for cholesterol
biosynthesis
- Subject to different kinds of metabolic
control
1. Sterol-dependent regulation of gene
expression
Sterol regulatory element-binding
protein-2 (SREBP-2)
- Transcription factor that controls the gene
expression for HMG CoA reductase
- Binds DNA at the cis-acting sterol
regulatory element (SRE) of the
reductase gene
SREBP an integral protein of the ER
membrane
- Associates with a second ER membrane
protein, SCAP (SREBP cleavage-acting
protein)

IPP and DPP condense to form 10carbon geranyl pyrophosphate (GPP)

When the sterol level in the cell


is low, the SREBP-SCAP complex
is sent out of the ER to the Golgi

Second molecule of IPP condenses


with GPP to form 15-carbon farnesyl
pyrophosphate

In the Golgi SREBP is


sequentially acted upon by 2
proteases, which generate a
soluble fragment that enters the
nucleus, binds the SRE and
functions as a transcription
factor

2 molecules of FPP combine, releasing


pyrophosphate, and are reduced
forming the 30-carbon compound
squalene

Increased synthesis of HMG


CoA reductase
Squalene is converted to the sterol
lanosterol by a sequence of reactions
catalyzed by ER-associated enzymes
that use O2 and NADPH. Hydroxylation
of squalene triggers the cyclization of
the strucure of lanosterol

The conversion of lanosterol to


cholesterol results to shortening of the
carbon chain from 30 to 27 carbons,
removal of 2 methyl groups at carbon
4, migration of double bond from
carbon 8 to carbon 5, and reduction of
double bond between carbon 24 and
carbon 25

Increased cholesterol synthesis

If sterols are
abundant, they bind
to SCAP at its sterolsensing domain
Binding of SCAP to
other ER membrane
proteins (insigs) is
induced
Retention of SCAPSREBP complex in
the ER

5. Inhibition by drugs
Statin drugs structural analogs of HMG
CoA
- Are (or are metabolized to) reversible,
competitive inhibitors of HMG CoA
reductase
- Used to decrease plasma cholesterol
levels in patients with
hypercholesterolemia
IV.

Prevent the
activation of SREBP

Down-regulation of
cholesterol
synthesis

2. Sterol-accelerated enzyme
degradation
Reductase a sterol-sensing integral
protein of the ER membrane
sterol levels in the cell reductase
binds to insig proteins ubiquitination
and proteasomal degradation of the
reductase

Degradation of Cholesterol
Intact sterol nucleus is
converted to bile acids and
bile salts

Excreted in the feces and by


secretion of cholesterol into
the bile

Transported to the intestine for


elimination

Some of the choleterol in the


intestine is modified by
bacteria before excretion

3. Sterol-independent
phosphorylation/dephosphorylation
AMP-activated protein kinase (AMPK)
+ phosphoprotein phosphatase
controls covalently the activity of CoA
reductase
Phosphorylated inactive enzyme
Dephosphorylated active enzyme
AMPK activated by AMP
ATP availability, cholesterol synthesis

Coprostanol, cholestanol, and


cholesterol make up the bulk of neutral
fecal sterols

4. Hormonal regulation
insulin and thyroxine, upregulation of
expression of the gene for HMG CoA
reductase
glucagon and glucocorticoids,
downregulation of expression of the gene
for HMG CoA reductase

V.
Bile Acids and Bile Salts
Bile watery mixture of organic and inorganic
compounds
- Can either:
Pass directly from the liver where it
is synthesized into the duodenum
through the common bile duct

Primary compounds made are


isomers coprostanol and
cholestanol - reduced
derivatives of cholesterool

Stored in the gallbladder when not


immediately needed for digestion
Phosphatidylcholine (lecithin) and bile
salts (conjugated bile acids)
quantitatively the most important organic
compounds of bile

A. Structure of bile acids


Bile acids contain 24 carbons with 2 or 3
OH groups and a side chain that terminates in
a carboxyl group
- Amphipathic (-OH groups are in
orientation- lie below the plane of the
rings) can act as emulsifying agents in
the intestine; help prepare dietary TAG
and other complex lipids for degradation
by pancreatic digestive enzymes
- Methyl groups are (lie above the plane of
the rings)
Carboxyl group pKa 6
- Not fully ionized at physiologic pH
B. Synthesis of bile acids

-OH groups are inserted at


specific positions on the
steroid structure

Double bond of cholesterol


B rings is reduced

Hydrocarbon chain is
shortened by 3 carbons,
introducing a carboxyl
group at the end of the
chain
Product: "Primary" bile
acids:
cholic acid (triol)
and chenodeoxycholic acid
(diol)

Rate-limiting step: introduction of OH at


carbon 7 of the steroid nucleus by
cholesterol-7- -hydroxylase
Cholesterol-7- -hydroxylase an ERassociated cytochrome P450 (CYP) enzyme
found only in liver
- Down-regulated by cholic acid

C. Synthesis of bile salts


- Bile acids 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 of the added
compound before they leave the liver
New structures:
Glycocholic acid
Glycochenodeoxycholic acids
Taurocholic
Taurochenodeoxycholic acids
Taurine endproduct of cysteine metabolism
3:1 ratio of glycine to taurine forming in the
bile
Addition of glycine or taurine
- Results in the presence of carboxyl group
with a lower pKa (from glycine) or a
sulfonate group (from taurine) both of
which are fully ionized (negatively
charged) at physiologic pH
Bile salts conjugated forms
- More effective detergents than bile acids
because of their enhanced amphipathic
nature; thus, only the conjugated forms
are found in the bile
- Provide the only significant mechanism for
cholesterol excretion, both as a metabolic
product of cholesterol and as a solubilizer
of cholesterol in bile
Exogenously supplied chenodeoxycholic
acid
- Treatment for individuals with genetic
deficiencies in the conversion of
cholesterol to bile acids

D. Action of intestinal flora on bile salts


Bacteria in the intestine
- Can remove glycine and taurine from bile
salts regenerating bile acids
- Convert some of the primary bile acids
into secondary bile acids by removing a
OH group, producing:
Deoxycholic acid from cholic acid
Lithocholic acid from
chenodeoxycholic acid
E. Enterohepatic circulation
- Bile salts secreted into the intestine are
efficiently reabsorbed (> 95%) and reused
- Liver converts both primary and
secondary bile acids into bile salts by
conjugation with glycine or taurine
secreted into the bile
Ileum via a Na+-bile salt cotransporter
where bile acids + bile salts is primarily
absorbed
Bile acids + bile salts 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
cotransporter
Bile acids hydrophobic
- Require a carrier in the portal blood
Albumin carries bile acids in a noncovalent
complex
Enterohepatic circulation
- Continuous process of secretion of bile
salts into the bile passage through the
duodenum where some are converted to
bile acids uptake in the ileum
subsequent return to the liver as a mixture
of bile acids and salts
Bile acid sequesterants (e.g.
cholestyramine) bind bile acids in the gut
- Prevent reabsorption of bile acids
promote excretion
- Used in the treatment of
hypercholesterolemia because the
removal of bile acids relieves the inhibition

on bile acid synthesis in the liver divert


additional cholesterol into that pathway
Dietary fiber also binds bile acids and
increases their excretion
F. Bile salt deficiency: cholelithiasis
- disruption of the simultaneous movement
of cholesterol from the liver into the bile
and secretion of phospholipid and bile
salts more cholesterol enters the bile
than can be solubilized by the bile salts
and phosphatidylcholine present
precipitation of cholesterol in the
gallbladder leading to cholesterol
gallstone disease
- typically caused by bile acids in the bile
which may result from:
gross malabsorption of bile acids
from the intestine (seen in patients
with severe ileal disease)
obstruction of biliary tract
interrupted enterohepatic
circulation
severe hepatic dysfunction
decreased synthesis of bile salts or
other abnormalities in bile
production
excessive feedback suppression of
bile acid synthesis due to
accelerated rate of recycling of bile
acids
- may also result from increased biliary
cholesterol excretion (seen with use of
fibrates)
Fibrates (e.g. gemfibrozil) derivatives of
fibric acid
- used to reduce TAG levels in blood through
up-regulation of fatty acid -oxidation
Laparoscopic cholecystectomy surgical
removal of gallbladder through a small
incision
- treatment of choice
Oral administration of chenodeoxycholic
acid
- for patients who are unable to undergo
surgery

supplement bodys supply of bile acids


gradual (months to years) dissolution of
gallstones

VI.
Plasma Lipoproteins
- Spherical macromolecular complexes of
lipids and specific proteins
(apolipoproteins or apoproteins)
Lipoprotein particles
- Include:
Chylomicrons (CM)
Very-low-density lipoproteins
Low-density lipoproteins
High-density lipoproteins
- Differ in lipid and protein composition,
size, density and site of origin
- Function both:
to keep their component lipids
soluble as they transport them in
the plasma
to provide an efficient mechanism
for transporting their lipid contents
to (and from) the tissues
- Humans experience a gradual deposition
of lipid (especially cholesterol) in tissues
potentially life-threatening occurrence
when the lipid deposition contributes to
plaque formation atherosclerosis
A. Composition of plasma lipoproteins
Lipoproteins neutral lipid core (TAG +
cholesteryl esters) surrounded by a shell of
amphipathic apolipoproteins, phospholipid
and nonesterified (free) cholesterol
- Constantly interchange lipids and
apolipoproteins with each other
Shell of amphipathic apolipoproteins,
phospholipid, and nonesterified cholesterol
(free)
- Oriented so that their polar portions are
exposed on the surface of the lipoprotein,
thus making the particle soluble in
aqueous solution
TAG and chlolesterol carried by
lipoproteins are obtained from:
Diet (exogenous source)
De novo synthesis (endogenous
source)

1. Size and density of lipoprotein


particles
Chylomicrons lipoprotein particles
lowest in density and largest in size
Contain the highest percentage of
lipid
Lowest percentage of protein
VLDLs and LDLs
Successively denser
Higher ratios of protein to lipid
HDL particles
Densest
Plasma lipoproteins
Can be separated on the basis of
their electrophoretic mobility or on the
basis of their density by
ultracentrifugation
2. Apolipoproteins
- Associated with lipoprotein particles
Functions:
Provide recognition sites for cellsurface receptors
Serve as activators or coenzymes for
enzymes involved in lipoprotein
metabolism
- Required as essential structural
components of the particles and cannot be
removed (particles cannot be produced
without them), whereas others are
transferred freely between lipoproteins
5 Major Classes
A through E
Subclasses
Apolipoprotein (or apo) A-I
Apo C-II

B. Metabolism of chylomicrons
Intestinal mucosal cells where
chylomicrons are assembled
Chylomicrons carry:
Dietary TAG

Cholesterol
Fat-soluble vitamins
Cholesteryl esters (plus additional
lipids made in these cells)
to the peripheral tissues
TAG account for close to 90% of lipids in a
chylomicron
1. Synthesis of apolipoproteins
Apolipoprotein B-48 unique to
chylomicrons
- Constitutes the N-terminal, 48% of the
protein coded for by the gene for apo B
Rough ER where synthesis of
apolipoprotein B-48 begins glycosylated as
it moves through the RER and Golgi
Apo B-100 synthesized by the liver
- Found in VLDL and LDL
- Represents the entire protein coded for by
the apo B gene
Nonsense codon created by
posttranscriptional editing of a cytosine to a
uracil in intestinal apo B-100 mRNA
- Allow translation of only 48% of mRNA
2. Assembly of chylomicrons
- Occurs before transition from the ER to the
Golgi, where the particles are packaged in
secretory vesicles fuse with the plasma
membrane releasing lipoproteins enter
the lymphatic system enter the blood
Smooth ER where enzymes involved in
TAG, cholesterol, and phospholipid synthesis
are located
Microsomal TAG transfer protein
required in assembly of apolipoproteins and
lipid into chylomicrons
- Loads apo B-48 with lipid
3. Modification of nascent chylomicron
particles
Nascent chylomicron particle released
by the intestinal mucosal cell
- Receives apolipoprotein E and C when it
reaches the plasma
Apolipoprotein E recognized by hepatic
receptors

Apolipoprotein C includes apo C-II


necessary for activation of lipoprotein
lipase
Lipoprotein lipase degrades the TAG
contained in the chylomicron
HDL source of these apolipoproteins
4. Degradation of TAG by lipoprotein
lipase
Lipoprotein lipase extracellular enzyme
that is anchored by heparin sulfate to the
capillary walls of most tissues, but
predominantly those of:
adipose tissue
cardiac muscle
skeletal muscle
- activated by apo C-II on circulating
lipoprotein particles
- hydrolyzes TAG contained in lipoprotein
particles to yield fatty acids and glycerol
Adult liver does not have lipoprotein
lipase
Hepatic lipase found on the surface of
endothelial cells of the liver
- plays some role in TAG degradation in CM
and VLDL
- particularly important in HDL metabolism
Fatty acids stored by the adipose or used
for energy by the muscle
- if not immediately taken up by a cell, LCFA
are transported by serum albumin until
their uptake does occur
Glycerol used by the liver in:
lipid synthesis
glycolysis
gluconeogenesis
Lipoprotein lipase deficiency or apo C-II
(Type 1 hyperlipoproteinemia or familial
lipoprotein lipase deficiency)
- dramatic accumulation of chylomicron-TAG
in the plasma (hypertriacylglycerolemia)
even in the fasted state
5. Regulation of lipoprotein lipase
activity
Insulin stimulate lipoprotein lipase
synthesis and transfer to the luminal surface
of the capillary (fed state)

Adipose enzyme has a large Km allows


removal of fatty acids from circulating
lipoprotein particles and their storage as TAG
only when plasma lipoprotein concentrations
are elevated

Chylomicron remnants bind to


lipoprotein receptors

Taken into the hepatocytes by


endocytosis
Heart muscle lipoprotein lipase has a
small Km
- Allows the heart continuing access to the
circulating fuel, even when plasma
lipoprotein concentrations are low
Cardiac muscle has the highest
concentration of lipoprotein lipase reflect
the use of fatty acids to provide much energy
needed for cardiac function
6. Formation of chylomicron remnants
- As the chylomicron circulates and more
than 90% of TAG in its core is degraded by
lipoprotein lipase, the particle size
and density
- C apoproteins but not apo E are returned
to HDL
Remnant rapidly removed from the
circulation by the liver cell membranes
contain lipoprotein receptors that recognize
apo E

Endocytosed vesicle fuses


with a lysosome

Apolipoproteins, cholesteryl
esters, and other components
of the remnant are
hydrolytically degraded

Amino acids, free cholesterol,


and fatty acids are released

Receptor is recycled

C. Metabolism of VLDL
VLDL produced in the liver
- Composed predominantly of endogenous
TAG (approximately 60%)
- Function: carry endogenous TAG from the
liver (site of synthesis) to the peripheral
tissues
Peripheral tissues where TAG is degraded
by lipoprotein lipase
Fatty liver (hepatic steatosis) occurs in
conditions in which there is an imbalance
between hepatic TAG synthesis and the
secretion of VLDL
- Characterized by:
Obesity
Uncontrolled diabetes mellitus
Chronic ethanol digestion
1. Release of VLDL

VLDL are secreted directly into the blood


by the liver as nascent VLDL particles
containing apo B-100 must obtain apo CII and apo E from circulating HDL
Chylomicrons
- Apo C-II is required for activation of
lipoprotein lipase
Abetalipoproteinemia rare
hypolipoproteinemia
- Caused by a defect in microsomal TAG
transfer protein (MTP) inability to load
apo B with lipid no VLDL or
chylomicrons are formed and TAG
accumulate in the liver and intestine

VLDL pass through the


circulation

TAG is degraded by
lipoprotein lipase

VLDL decrease in size and


become denser

Surface components, including


the C and E apoproteins, are
returned to HDL, but the
particles retain apo B-100

Some TAG are transferred from


VLDL to HDL in an exchange
reaction that concomitantly
transfers some cholesteryl
esters from HDL to VLDL
(accomplished by cholesteryl
ester transfer protein or CETP)

2. Modification of circulating VLDL

3. Production of LDL from VLDL in the


plasma
- VLDL is converted in the plasma to LDL
Intermediate-density lipoproteins (IDL)
or VLDL remnants observed during this
transition
IDL can also be taken up by cells through
receptor-mediated endocytosis that uses apo
E as the ligand
Apo E normally present in 3 isoforms:
E-2 binds poorly to receptors
E-3
E-4
Patients homozygotic for apo E-2 are deficient
in the clearance of chylomicron remnants and
IDL have Type III hyperlipoproteinemia
(familial dysbetalipoproteinemia or
broad beta disease) with
hypercholesterolemia and premature
atherosclerosis
- E-4 isoform confers increased
susceptibility to and decreased age of
onset of late-onset Alzheimer disease,
doubling lifetime risk
D. Metabolism of LDL
LDL contain much less TAG than their VLDL
predecessors
- Have a high concentration of cholesterol
and cholesteryl esters
1. Receptor-mediated endocytosis
Primary function of LDL particles: provide
cholesterol to the peripheral tissues (or return
it to the liver) by binding to cell surface
membrane LDL receptors that recognize apo
B-100 (but not apo B-48) these receptors
can also bind apo E; they are also known as
apo B-100/apo E receptors

LDL receptors are negatively charged


glycoproteins that are clustered in pits on
cell membranes. The cytosolic side of the
pit is coated with the protein clathrin,
which stabilizes the shape of the pit

After binding, the LDL-receptor complex is


internalized by endocytosis

Type II hyperlipidemia (familial


hypercholesterolemia, FH) and premature
atherosclerosis
- deficiency of functional LDL receptors
- plasma LDL and plasma cholesterol
FH can also be caused by:
protease activity that degrades the
receptor
Defects in apo B-100 that reduce its
binding to the receptor
CURL compartment for uncoupling of receptor
and ligand
- Where receptors migrate to
Wolman disease
- Storage disease caused by rare autosomal
recessive deficiencies in the ability to
hydrolyze lysosomal cholesteryl esters

The vesicle containing LDL loses its


clathrin coat and fuses with other similar
vesicles, forming larger vesicles
(endosomes)

THe pH of the endosome falls due to the


proton-pumping activity of endosomal
ATPase - allows separation of LDL from
its receptor

Receptor 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
transferred to lysosomes and degraded by
lysosomal acid hydrolases, releasing
free cholesterol, amino acids, fatty acids,
and phospholipids. These compounds are
reutralized by the cell.

Niemann-Pick disease, Type C


- Inability to transport unesterified
cholesterol out of the lysosome
2. Effect of endocytosed cholesterol on
cellular cholesterol homeostasis
a. HMG CoA reductase is inhibited by
cholesterol;
de novo cholesterol synthesis
b. synthesis of new LDL receptor protein
by expression of LDL receptor gene
limited entry of LDL cholesterol into cells
c. If the cholesterol is not required
immediately for some structural or
synthetic purpose, it is esterified by acyl
CoA: cholesterol acyltransferase
(ACAT)
SRE and SREBP (SREBP-2) involved in the
regulation of LDL receptor gene
ACAT transfers fatty acid from fatty acyl
CoA derivative to cholesterol; Product:
cholesteryl ester that can be stored in the cell
- Activity is enhanced in the presence of
increased intracellular cholesterol
3. Uptake of chemically modified LDL by
macrophage scavenger
Macrophages possess high levels of
scavenger receptor activity

Scavenger receptor class A (SR-A)


- Can bind a broad range of ligands
- Mediate endocytosis of chemically
modified LDL in which the lipid
components of apo B have been oxidized
- Not down-regulated in response to
intracellular cholesterol
Cholesteryl esters accumulate in
macrophages
- Cause transformation of macrophages into
foam cells participate in the formation
of atherosclerotic plaque
E. Metabolism of HDL
HDL comprise of heterogeneous family of
lipoproteins with a complex metabolism
HDL particles formed in blood by the
addition of lipid
to apo A-1
Apo A-1 apolipoprotein made by the liver
and intestine and secreted into blood
- Accounts for about 70% of the apoproteins
in HDL
Functions of HDL
1. HDL is a reservoir of apolipoproteins
HDL particles serve as circulating reservoir
of apo C-II
Apo C-II apolipoprotein that is transferred
to VLDL and chylomicrons
- Activator of lipoprotein lipase
Apo E apolipoprotein required for the
receptor-mediated endocytosis of IDLs and
chylomicron remnants
2. HDL uptake of unesterified
cholesterol
Nascent HDL disk-shaped particles
containing primarily phospholipid (largely
phosphatidylcholine) and apolipoproteins A, C,
and E
- Take up cholesterol from non-hepatic
(peripheral) tissues and return it to the
liver as cholesteryl esters
HDL particles excellent acceptors of
unesterified cholesterol as a result of their
high concentration of phospholipids, which
are important solubilizers of cholesterol
3. Esterification of cholesterol

Cholesterol when taken up by HDL, it is


immediately esterified by the plasma enzyme
lecithin:cholesterol acyltransferase
(LCAT or PCAT; P = Phosphatidylcholine)
LCAT synthesized by the liver
- Binds to nascent HDL
- Activated by: Apo A-I
- Transfers fatty acid from carbon 2 of
phosphatidylcholine to cholesterol
Product: hydrophobic cholesteryl ester
(sequestered in the core of HDL) and
lysophosphatidylcholine (binds to albumin)
Esterification maintains cholesterol
concentration gradient allow continued
efflux of cholesterol to HDL
Discoidal nascent HDL accumulates
cholesteryl esters
- First becomes a spherical, relatively
cholesteryl ester-poor HDL3 cholesteryl
ester-rich HDL2 particle that carries these
esters to the liver
Cholesterol ester transfer protein (CETP)
moves some of the cholesteryl esters from
HDL to VLDL in exchange for TAG relieve
product inhibition of LCAT
Because VLDL are catabolized to LDL,
cholesteryl esters are ultimately taken up by
the liver
4. Reverse cholesterol transport
- Involves
efflux of cholesterol from
peripheral cells to HDL mediated,
at least in part, by the transport
protein ABCA1
esterification of cholesterol by
LCAT
binding of cholesteryl ester-rich
HDL (HDL2) to liver and
steroidogenic cells
selective transfer of the cholesteryl
esters into these cells
release of lipid-depleted HDL
(HDL3)
Key component of cholesterol
homeostasis:

Selective transfer of cholesterol from


peripheral cells to HDL and from HDL to
the liver for bile acid synthesis or disposal
via the bile and to steroidogenic cells for
hormone synthesis
Basis for
inverse relationship seen between
plasma HDL concentration and
atherosclerosis
HDLs designation as the good
cholesterol carrier

Tangier disease very rare deficiency of


ABCA1
- Characterized by virtual absence of HDL
particles due to degradation of lipid-poor
apo A-1
SR-B1 (scavenger receptor class B type
1)
- Cell-surface receptor
- - mediates the uptake of cholesteryl esters
by the liver
- Binds HDL
Hepatic lipase can degrade both TAG and
phospholipids
- Participates in the conversion of HDL2 to
HDL3
ABCA1 an ATP-binding cassette (ABC)
protein
ABC proteins use energy from ATP
hydrolysis to transport materials, including
lipids, in and out of cells and across
intracellular compartments
Defects in specific ABC proteins result
in:
X-linked adrenoleukodystrophy
Respiratory distress syndrome due to
decreased surfactant secretion
Cystic fibrosis
F. Role of lipoprotein (a) in heart
disease
Lipoprotein (a) or Lp (a)
- Particle, when present in large quantities
in the plasma, is associated with an
increased risk of coronary heart disease

Nearly identical in structure to an LDL


particle
Distinguishing feature: presence of
additional apolipoprotein molecule (apo
(a)) that is covalently linked at a single
site to apo B-100

Circulating levels of Lp(a) are determined


primarily by genetics
Diet may play some role as trans fatty acids
have been shown to Lp(a)
Estrogen - both LDL and Lp(a)
Apo(a) structurally homologous to
plasminogen
Plasminogen precursor of blood protease
whose target is fibrin
Fibrin main protein component of blood
clots
Lp(a) slows down the breakdown of blood
clots that trigger heart attacks because it
competes with plasminogen for binding to
fibrin
Niacin reduces Lp(a) and raises HDL
VII.
Steroid Hormones
Cholesterol precursor of all classes of
steroid hormones:
Glucocorticoids e.g. cortisol
Mineralocorticoids e.g. aldosterone
Sex hormones e.g. androgens,
estrogens, and progestins
Corticosteroids collective term for
glucocorticoids and mineralocorticoids
Adrenal cortex where synthesis and
secretion of cortisol, aldosterone, and
androgens occur
Ovaries and placenta where synthesis and
secretion of estrogens and progestins occur
Testes where synthesis and secretion of
testosterone occurs
Steroid hormones transported by the
blood from their sites of synthesis to their
target organs
- Must be complexed with a plasma protein
because of their hydrophobicity

Plasma albumin can act as a nonspecific


carrier
- carry aldosterone
Specific steroid-carrier plasma proteins
- bind steroid hormones more tightly than
does albumin
E.g. corticosteroid-binding globulin
(transcortin)
- responsible for transporting cortisol
A. Synthesis of steroid hormones
- Involves shortening hydrocarbon chain of
cholesterol and hydroxylation of the
steroid nucleus
Initial and rate-limiting reaction
converts cholesterol to the 21-carbon
pregnenolone
Cholesterol side-chain cleavage enzyme
complex (desomolase, P450scc) catalyzes
the conversion of cholesterol to 21-carbon
pregnenolone
- A cytochrome P450 (CYP) mixed function
oxidase of the inner mitochondrial
membrane
NADPH and O2 required for reaction
Cholesterol substrate can be newly
synthesized, taken up from lipoproteins, or
released from cholesteryl esters stored in the
cytosol of steroidogenic tissues
Important control point:
- Movement of cholesterol into mitochondria
mediated by StAR (steroidogenic acute
regulatory protein)
Pregnenolone parent compound for all
steroid hormones
- Oxidized and then isomerized to
progesterone which is further modified to
the other steroid hormones by
hydroxylation reactions that occur in the
ER and mitochondria
Enzymes primarily are CYP proteins
- A defect in the activity or amount of an
enzyme in this pathway can lead to a
deficiency in the synthesis of hormones
beyond the affected step and to an excess
in the hormones or metabolites before
that step
Congenital adrenal hyperplasias
collective term for enzyme deficiencies

Addison disease due to autoimmune


destruction of the adrenal cortex
- Characterized by adrenocortical
insufficiency
B. Secretion of adrenal cortical steroid
hormones
Steroid hormones secreted on demand
from their tissues of origin in response to
hormonal signals
Corticosteroids and androgens made in
different regions of the adrenal cortex
- Secreted into blood in response to
different signals
1. Cortisol
Middle layer (zona fasciculate) of the
adrenal cortex where cortisol is produced
controlled by the hypothalamus to which
pituitary gland is attached
Corticotropin-releasing hormone (CRH)
- Produced by the hypothalamus
- Travels through capillaries to the anterior
lobe of the pituitary in response to severe
stress (e.g. infection)
- Induces production and secretion of
adrenocorticotropic hormone (ACTH) in the
pituitary
Polypeptide ACTH stress hormone
- Stimulates adrenal cortex to synthesize
and secrete the glucocorticoid cortisol
Cortisol allows the body to respond to
stress through its effects on intermediary
metabolism (e.g. increased gluconeogenesis)
and inflammatory and immune responses -
cortisol, release of CRH and ACTH is inhibited
ACTH binds to a membrane G-protein
coupled receptor results in cAMP production
and activation of protein kinase A
PKA phosphorylates the esterase that
converts cholesteryl ester to cholesterol and
stimulates synthesis of StAR protein
2. Aldosterone

Primary effect on kidney tubules:


stimulates sodium uptake and potassium
excretion
- BP
Outer layer (zona glomerulosa) of the
adrenal cortex where aldosterone is
produced induced by plasma Na+/K+ ratio
and by angiotensin II
Angiotensin II an octapeptide
- Produced from angiotensin I (decapeptide)
by angiotensin-converting enzyme (ACE)
found predominantly in the lungs, but
which is also distributed widely in the
body
- Binds to cell-surface receptors
- Effects are mediated through the
phosphatidylinositol 4,5-bisphosphate
(PIP2) pathway and not by cAMP
Inhibitors of ACE are used to treat renindependent hypertension
Angiotensin I produced in the blood by
cleavage of an inactive precursor
(angiotensin) secreted by the liver
- Cleavage is accomplished by the enzyme
renin, made and secreted by the kidney
3. Androgens
Inner (zona reticularis) and middle layers
of the adrenal cortex produce androgens,
primarily dehydroepiandrosterone and
androstenedione
Adrenal androgens weak
- Converted in peripheral tissues to
testosterone and to estrogens
Testosterone stronger androgen
Estrogens derived from androstenedione
and testosterone by aromatase (CYP19)
Aromatase inhibitors used in the
treatment of estrogen-responsive breast
cancer in post-menopausal women
C. Secretion of steroid hormones from
gonads
Testes and ovaries synthesize hormones
necessary for sexual differentiation and
reproduction
Gonadotropin-releasing hormone single
hypothalamic-releasing factor

Stimulates the anterior pituitary to


release the:
Glycoproteins
Luteinizing hormone (LH)
stimulates the testes to produce
testosterone and the ovaries to
produce estrogens and
progesterone
Follicle-stimulating hormone
(FSH) regulates the growth of
ovarian follicles and stimulates
testicular spermatogenesis
LH and FSH bind to surface receptors
- Cause an increase in cAMP

D. Mechanism of steroid hormone action


HRE found in the promoter (or an enhancer
element) for genes that respond to a specific
steroid hormone ensure coordinated
regulation of these genes
Hormone-receptor complexes can also
inhibit transcription in association with
corepressors
Binding of a hormone to its receptor causes a
conformational change in the receptor that
uncovers its DNA-binding domain allow
complex to interact through a Zn-finger motif
with the appropriate sequence on the DNA

Each steroid hormone


diffuses across the plasma
membrane of its target cell
and binds to a specific
cytosolic or nuclear receptor

Receptor-ligand complexes
accumulate in the nucleus

Receptor-ligand complexes
dimerize

Bind to specific regulatory


DNA sequences (hromoneresponse elements, HRE) in
association with coactivator
proteins

Promoter activation and


increased transcription of
targeted genes

Receptors for steroid hormones, thyroid


hormone, retinoic acid, and 1,25dihydroxycholecalciferol (vitamin D)
members of a superfamily of structurally
related gene regulators that function in a
similar way
E. Further metabolism of steroid
hormones
Steroid hormones generally converted
into inactive metabolic excretion products in
the liver
Reactions include:
Reduction of unsaturated bonds
Introduction of additional hydroxyl
groups
Resulting structure: made more soluble by
conjugation with glucuronic acid or sulfate
(from 3-phosphoadenosyl5phosphosulfate)
Approximately 20-30% of these metabolites
are secreted into the bile and then excreted in
the feces
The remainder are released into the blood and
filtered from the plasma in the kidney,
passing into the urine
Conjugated metabolites are fairly watersoluble and do not need protein carriers