cholesterol structure

• most plasma cholesterol is in

the esterified form (not found
in cells or membranes)

• cholesterol functions in all

membranes (drives formation of
lipid microdomains)

• cholesterol is the precursor for

steroid hormones

• note 4 fused rings, single dbl

bond, single hydroxyl, acyl
chain at C17
1

Cholesterol FAQs

• Cholesterol is the second most abundant

fraction in blood besides glucose.

• All the carbon atoms in cholesterol come from

Acetyl-CoA.

• Energy for synthesis comes from hydrolysis of

thioester bonds of acetyl CoA and ATP
hydrolysis.

• Synthesis occurs in the cytoplasm, with key

enzymes found in the membrane of the ER.

Cholesterol
promotes the
“liquid-ordered”
phase of
membranes
• cholesterol has limited
flexibility and is amphipathic.
• it stiffens the membrane and
regulates permeability.
• it can interact with and affect
the structure of integral
membrane proteins (e.g. lipid
rafts)
3
Friday, October 15, 2010
 S.R. Wassall, W. Stillwell / Chemistry and Physics of Lipids 153 (2008) 57–63

Cholesterol is less soluble in

(artificial) membranes high in
unsaturated acyl chains
4

Journal of Lipid Research,Vol. 44, 655-667, April 2003

5

Lynen F. DerWegvonder. ‘‘AktiviertenEssigsa ̈use’’ zuden

terpenen und den fettsa ̈ uren. Les Prix Nobel. Stockholm:
Norstedt & Sons, 1965: 205–45.
1964 Nobel Prize for discovery of biosynthesis of cholesterol Zetterström

Figure 6 Biosynthesis of terpenes. From Lynen (4).

to mevalonic acid may be the homeostatic control mecha-

nism in the biosynthesis of cholesterol, a hypothesis, which
is now known to be correct.

6
References
Friday, October 15, 2010 1. Ružička L. The isoprene rule and the biogenesis of terpenic
compounds. Experientia 1953; 9: 35–7.
2. Robinson R. J Chem Soc Ind Lond 1934; 53: 1062–3.
3. Channon HJ. The biological significance of the unsaponifiable
matter of oils: experiments with the unsaturated hydrocarbon,
Figure 7 Biosynthesis of fatty acids according to a hypothetic structure of a
 Cholesterol synthesis initially
follows that of ketone bodies

3 cytoplasmic acetyl
CoA molecules are
sequentially
condensed to form
HMG CoA (6 carbons)

cytoplasmic
HMG-CoA Synthase

mitochondrial
HMG-CoA Synthase

Cholesterol biosynthesis includes over 30 enzymatic steps

that occur in the cytoplasm and outer membrane of the ER
8

Mito_HMG- Cyto_HMG-
CoA CoA
Synthase Synthase

9
Friday, October 15, 2010
 10

11

12
Friday, October 15, 2010
 13

Biochemistry, Vol. 39, No. 8, 2000

Beginning with squalene, sterol-

carrier-protein keeps the remaining
intermediates soluble
14

Squalene epoxide is converted to

lanosterol in concerted fashion

15
Friday, October 15, 2010
 16

The committed step of de novo

cholesterol biosynthesis is
catalyzed by HMG CoA reductase
• The reduction of HMG CoA by
HMG CoA reductase results in
the oxidation of two NADPH
and results in mevalonate.

• HMG CoA reductase is a

membrane protein of the ER:
catalytic domain projects into
the cytoplasm.

• Target of statin drugs

17

The committed step of de novo

cholesterol biosynthesis is
catalyzed by HMG CoA reductase

18
Friday, October 15, 2010
 HMG-CoA-reductase forms a
homotetramer and utilizes NADPH
and CoA cofactors
19

Cell Research (2008) 18:609–621

HMG-CoA-r has a cytoplasmic

catalytic domain and a membrane
sterol sensing domain
20

HMG-CoA-reductase utilizes two

NADPH for each HMG-CoA

1. NADPH reduces HMG carbon.

2. His donates a proton and cleaves CoA from HMG.
3. Second NADPH protonates HMG carbonyl to form
Mevalonate.
21
Friday, October 15, 2010
 Fine control of cholesterol
homeostasis

• Inhibitory phosphorylation of HMG-CoA-r at

SER872 (e.g. by AMP-dependent protein kinase)
and regulated by hormones.
22

Coarse control of cholesterol

homeostasis
QDWH SDWKZD\
SCAP SREBP Nucleus
egradation of Reg E+/+
eral excellent ER SRE

,QVLJ E+/+
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hat mediate Sterols

CoA reduc-

E+/+ E+/+
terol-acceler- Golgi
d from com-
(the SREBP Lumen
S1P S2P
ductase, Scap
ic N-terminal 1.Figure Regulation of the transcription of HMG-CoA-r
3 Model for sterol-regulated Scap-SREBP pathway. SCAP
and a hydro- (and others) by SCAP/SREBP (sterol-response-
LVDVHQVRURIVWHUROVDQGDQHVFRUWRI65(%3V,QVWHUROGHSOHWHG
e cytosol [24]. cells, Scap facilitates export of SREBPs from the ER to the Golgi
element-binding-protein)
a constitutive apparatus, where two proteases, Site-1 protease (S1P) and Site-2 23
s required for SURWHDVH 63 DFW WR UHOHDVH WKH WUDQVFULSWLRQDOO\ DFWLYH 1
om the ER to WHUPLQDOE+/+=LSGRPDLQRI65(%3VIURPWKHPHPEUDQH7KH
UHOHDVHGE+/+=LSGRPDLQPLJUDWHVLQWRWKHQXFOHXVDQGELQGVWRD
n arrival in the
sterol response element (SRE) in the enhancer/promoter region of
s (designated
vely to release Coarse control of cholesterol
WDUJHWJHQHVDFWLYDWLQJWKHLUWUDQVFULSWLRQ$FFXPXODWLRQRIVWHUROV
LQ(5PHPEUDQHVWULJJHUVELQGLQJRI6FDSWRRQHRIWZRUHWHQWLRQ
npg Sterol-accelerated degradation of HMG CoA reductase

o the cytosol homeostasis

618
SURWHLQVFDOOHG,QVLJVZKLFKEORFNVLQFRUSRUDWLRQRI6FDS65(%3
then migrate FRPSOH[HVLQWR(5WUDQVSRUWYHVLFOHV$VDUHVXOW65(%3VQR
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te target gene ORQJHUWUDQVORFDWHWRWKH*ROJLDSSDUDWXVWKHE+/+=LSGRPDLQ
8EF
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sis and uptake FDQQRWEHUHOHDVHGIURPWKHPHPEUDQHDQGWUDQVFULSWLRQRIDOO
gp78 reductase
ELQGLQJ
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targetERgenes declines.
n of sterols in Lumen
,QVLJ
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on of SREBPs
from the ER; 8ELTXLWLQDWLRQ

s and choles- tase. This effort led to the following 8IG observations, which
hibition of ER considered
Degradation together divulge the action of at least one of
9&3
9&3

terol-induced the Insig proteins 8EF in sterol-accelerated8EFdegradation of

gp78 gp78
called Insig-1 reductase. First, when overexpressed Extraction by transfection in
es a cytosolic Chinese Proteasome
hamster ovary (CHO) cells, reductase cannot be
oteins, which degraded when the cells are treated with sterols [37]. Co-
OH
Geranylgeraniol

at deliver ER-Figure 5expression of Insig-1

Pathway for sterol-accelerated degradationrestores sterol-accelerated
of HMG CoA reductase. degrada-
Accumulation of 25-hydroxycholesterol, lanosterol,

2.tion INSIG proteinsuggesting

of reductase, senses sterol concentration
the saturation of endogenous and
RUGLK\GURODQRVWHUROLQ(5PHPEUDQHVWULJJHUVELQGLQJRIWKHUHGXFWDVHWR,QVLJV$VXEVHWRI,QVLJVLVDVVRFLDWHGZLWK
sig binding isWKHPHPEUDQHDQFKRUHGXELTXLWLQOLJDVHJSZKLFKELQGVWKH(8EFDQG9&3DQ$73DVHWKDWSOD\VDUROHLQH[WUDFWLRQRI
XELTXLWLQDWHGSURWHLQVIURP(5PHPEUDQHV7KURXJKWKHDFWLRQRIJSDQG8EFUHGXFWDVHEHFRPHVXELTXLWLQDWHGZKLFKWULJ-
e domain thatJHUVLWVH[WUDFWLRQIURPWKHPHPEUDQHE\9&3DQGVXEVHTXHQWGHOLYHU\WRSURWHDVRPHVIRUGHJUDGDWLRQ7KHSRVWXELTXLWLQDWLRQ
Insigs
regulates by the ubiquitin-mediated
overexpressed reductase.protein Second, reduction
VWHSLVSRVWXODWHGWREHHQKDQFHGE\JHUDQ\OJHUDQLROWKURXJKDQXQGH¿QHGPHFKDQLVPWKDWPD\LQYROYHDJHUDQ\OJHUDQ\ODWHG
0]. A similarSURWHLQVXFKDVRQHRIWKH5DESURWHLQV
of both Insig-1 and Insig-2 by RNA interference (RNAi)
degradation of HMG-CoA-Reductase
n at least four abolishes sterol-accelerated degradation of endogenous
g the Niemann studies thatreductase
directly focus on[38].reductaseThird,
degradationmutant
are CHO
underlying cellsforlacking
mechanisms both
sterol-accelerated, ERAD of
24
ductase),Friday,
all of October
stability toInsigs
overall regulation 15, in2010
required in order to determine the contribution of protein
are impervious
of reductase mice to
reductase may have implications for degradation of other
sterol-stimulated
in vivo degradation
FOLQLFDOO\ LPSRUWDQW SURWHLQV of¿EURVLV
VXFK DV WKH F\VWLF
sterols. Thus, under various physiologic conditions, such as hypoxia.
reductase
The significance as well as
of Insig-mediated regulation of
transmembrane conductance regulator (CFTR). Thus,
sterol-mediated inhibition
further excitement will undoubtedlyofensue
SREBP once questions
nsing domain reductase in processing [39]. homeostasis is posed in this review begin to become clear.
maintenance of cholesterol
 Summary of cholesterol homeostasis

Transcription HMG-CoA-r Degradation

P
HMG-CoA-r

[sterol] compensatory regulation

phosphorylation
INSIG-mediated degradation
dephosphorylation
SREB-mediated transcription

25

SCIENCE VOL 292 11 MAY 2001

Statins are competitive inhibitors

of HMG-CoA-reductase

26

HMG
CoA
NADP+

monomer-1
monomer-2

Statin binding blocks HMG-CoA

binding site
27
Friday, October 15, 2010
 Statin binding blocks the
Coenzyme-A binding pocket
28

Plant sterols Ezetimibe

• Reduces amount of • Reduces amount of

circulating cholesterol circulating cholesterol
(inhibits absorption)
• not compensated by
increased de novo • compensated for by
synthesis of increased de novo
cholesterol synthesis

29

Plant sterol margarines (Benecol, sitosterol) act

through a different mechanism than Ezetimibe

• Recent evidence points to enhanced degradation

of HMG-CoA-r.
30
Friday, October 15, 2010
 Cholesterol is modified by cytochrome p450
enzymes to form steroid hormones

31

Degradation of Cholesterol

• Unlike fatty acids, sterols cannot be used as an

energy source

• The sterol ring nucleus is eliminated from the

body by conversion to bile acids and bile salts.

32

Degradation of Cholesterol

• The theme is for cholesterol to be converted to a

relatively soluble amphipathic molecule.

• As a bonus, these molecules are used as

emulsifying agents during digestion.
33
Friday, October 15, 2010
 Lipoprotein particles transport lipids.

Each lipoprotein recruits a different

set of lipid transfer proteins
34

LDL delivers cholesterol directly to

the interior of cells
35

HDL are secreted ‘empty’ and scavenge

cholesterol via Apo-A associated LCAT (lecithin-
cholesteral acyltransferase) activity

36
Friday, October 15, 2010
 Lecithin-cholesteryl-acyltransferase (LCAT) is
recruited to HDL via Apolipoprotein A1(Apo A)

LCAT helps to sequester cholesterol (amphipathic)

by esterifying it to a (hydrophobic) fatty acid
37

Biophysical Journal, 88:548-556, 2005

apolipoprotein A1 homodimer forms

an α-helical belt to capture lipids 38

Friday, October 15, 2010