Research: Science and Education
edited by
Concepts in Biochemistry
Cholesterol Biosynthesis: Lanosterol to Cholesterol
John M. Risley
Department of Chemistry, The University of North Carolina at Charlotte, Charlotte, NC 28223-0001;
jmrisley@email.uncc.edu
Cholesterol is an important molecule. It is important
biochemically because it maintains membrane fluidity,
microdomain structure, and permeability, and it is a precursor
to steroid hormones, bile acids, vitamin D, and lipoproteins.
In addition to these well-recognized functions, cholesterol has
been shown recently to have an essential role in mammalian
embryonic development (1). It is important medically because
it is correlated with a number of diseases, as, for example,
atherosclerosis (2), cholestasis (broad and variable impairment
of bile secretion) (3), hypercholesterolemia (4 ), cholesterol
gallstone disease (5), and Nieman–Pick type C disease (6 ),
on which major research efforts and monies are expended
annually. It is important commercially for the pharmaceutical
industry in terms of drug development, and for the food
industry in terms of “healthy foods” that are advertised either
to be “cholesterol-free” or to reduce cholesterol. For example,
Zocor (simvastatin) has been advertised to the general public
by Merck & Co. as “an effective medicine that along with
diet and exercise can significantly lower total cholesterol” (7).
And, above the nutrition facts on a box of Kellogg’s Corn
Flakes the cereal is touted as a “cholesterol free food”, and
on the front of a box of General Mills Cheerios it reads “As
part of a heart-healthy diet, the soluble fiber in Cheerios May
Reduce Your Cholesterol!” Each cereal “Meets American Heart
Association food criteria for saturated fat and cholesterol for
healthy people over age 2”.
While exogenous cholesterol is considered a major cause
for many of the health problems, cholesterol biosynthesis in
vivo (endogenous cholesterol) is a target in reducing overall
cholesterol in humans (as, for example, by lovastatin, which
inhibits HMG-CoA reductase) (2). Considering the significance
and interest in cholesterol, it is interesting that discussions
of cholesterol biosynthesis in general biochemistry textbooks
are restricted to the synthesis of lanosterol from acetate, which
is usually discussed in detail. The conversion of lanosterol to
cholesterol is most often simply indicated, without elaboration,
as a multistep process. The only textbook I know of that
presents a pathway of lanosterol to cholesterol is that by Voet
and Voet (8), after Rilling and Chayet in 1985 (9). The
Biochemical Pathways chart distributed by Roche Molecular
Biochemicals (10) gives an abbreviated pathway showing
major intermediates, but in general without elaboration; the
chart is used by ExPASy with links to a few of the enzymes
(11). Map 100 in the Kyoto Encyclopedia of Genes and Genomes
(KEGG ) (12) outlines the pathway, naming major inter-
mediates with links to a few of the enzymes, but without
elaboration. This situation is unsatisfactory for numerous
students, especially those more medically oriented.
The research literature on cholesterol biosynthesis is vo-
luminous. A search of “cholesterol biosynthesis” on PubMed
JChemEd.chem.wisc.edu • Vol. 79 No. 3 March 2002 • Journal of Chemical Education
William M. Scovell
Bowling Green State University
Bowling Green, OH 43403
at the National Library of Medicine (13) yielded more than
10,000 citations, while a search of “cholesterol biosynthesis,
review” resulted in more than 1000 citations. A significant
proportion of the latter are reviews of drug effects on choles-
terol biosynthesis. However, I am not aware of a recent review
of the pathway from lanosterol to cholesterol that describes
in detail the reactions along the line of ref 9. I did find an
excellent review published in 1997 that summarizes the cloning
of cDNAs or genes encoding the enzymes of sterol biosyn-
thesis for fungi, mammals, and higher plants (14), including brief
descriptions of the enzyme reactions for each of the pathways
to ergosterol, cholesterol, stigmasterol, and campesterol. Ad-
vances have been made since the publication of the review
article. For example, the genes for C3 sterol dehydrogenase
(C4 decarboxylase) (15) and 3-keto reductase (16 ) have been
cloned, completing the elucidation of the pathway of ergos-
terol biosynthesis in the yeast Saccharomyces cerevisiae.
With no recent review from which to teach the conversion
of lanosterol to cholesterol, I have used the outline presented
in 1985 (9), with supplements obtained in my readings from
the research literature describing subsequent information on the
enzymes in this pathway. This paper presents the material
that I use in teaching this topic. It is divided into five sections:
(i) general overview of the pathway, (ii) step-by-step synthesis,
(iii) cholesterol biosynthesis in perspective, (iv) cholesterol
homeostasis, and (v) inherited diseases of cholesterol biosynthe-
sis. I do not present a comprehensive review of the literature in
this area, but I have included recent relevant references from
which a more comprehensive study may be pursued if desired.
Overview of the Pathway
The anabolic pathway for the conversion of lanosterol
to cholesterol is shown in Figure 1. The numbering scheme
for lanosterol is shown in Figure 2.
The biosynthesis of cholesterol from lanosterol is a 19-
step process. It requires nine different enzymes; two enzymes
catalyze multiple steps and three are utilized two times in
the pathway. Three methyl groups in lanosterol (Fig. 2: C30,
C31, C32) are oxidized and removed as formic acid and two
carbon dioxide molecules. Two mechanisms are utilized in
these reactions: a cytochrome P450 and formation of a β-
ketoacid. The other reactions in the pathway are five reduction
reactions using NAD(P)H, one dehydrogenation (desaturation)
reaction using NADH/O2, and one isomerization. Two mecha-
nisms are utilized in the reduction reactions: two ketoreductions
and three olefinic reductions involving carbocation intermedi-
ates. A radical mechanism is utilized in the dehydrogenation
(desaturation) reaction, and formation of a carbocation inter-
mediate is utilized in the isomerization reaction.
377
 Research: Science and Education

Figure 1. The pathway for the 19-step conversion of lanosterol to cholesterol.

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 Research: Science and Education

Step-by-Step Synthesis 22 24
27
21
18 20 23 25
In the following discussion of the 19-step reaction, I give
the various names used in the literature for the enzyme, sub- 19 11
12
13
17
16
26

strates, and products in each step. 9 14 15

1 8
10 α
Oxidative Demethylation of 1 to 5 β 3
2
5 32
7
6
The oxidative demethylation of lanosterol (4,4′,14α-tri- HO β
4
α
methyl-5α-cholesta-8(9),24-dien-3β-ol) (1) to cholestatriene 31 30
(4,4′-dimethyl-5α -cholesta-8(9),14,24-trien-3β -ol; 4,4′-
dimethylcholesta-8,14,24-trien-3β-ol) (5) is catalyzed by lanos- Figure 2. The numbering scheme for lanosterol.
terol 14α-methyl demethylase cytochrome P450 (lanosterol
14α-methyl demethylase; 14α-methyl demethylase; lanosterol
14α-demethylase) (17–22). The 14α-methyl group, which
protrudes into the α face of the sterol ring system, is the first
group to be removed; its removal is essential for membrane and
regulatory functions of the resulting sterols (14). The enzyme, Cl
a cytochrome P450, requires the C3-hydroxy group, 14α- N
methyl group, and ∆8(9) bond for binding and catalysis (22); it N CH2 CH O CH2 Cl
also requires three NADPH coenzymes and three O2 molecules. Cl
Upon substrate binding the heme shifts from a low-spin to a
high-spin state (22). miconazole
The enzyme catalyzes three successive monooxygenation Cl
reactions: (i) lanosterol (1) to 3β-hydroxylanost-8-en-32-ol
(lanost-8-en-3β,32-diol) (2), (ii) 3β-hydroxylanost-8-en-32-ol
(2) to 3β-hydroxylanost-8-en-32-al (3), and (iii) 3β-hydroxy- O N N COCH3
lanost-8-en-32-al (3) to 14α-formyloxy-lanost-8-en-3β-ol (4) O
N
to cholestatriene (5). The third oxidation has been proposed N CH2 C O
to proceed by peroxy attack, a Baeyer–Villiger rearrangement, Cl
and deformylation by a base rearrangement (17), or by peroxy ketoconazole
attack, homolytic cleavage, deformylation by fragmentation,
and disproportionation (20). (Deformylation by fragmentation
has been termed a radicalar mechanism [14].) The human cDNA Cl
clone has been reported (23) and a model of the Candida Figure 3. The structures for two inhibitors of lanosterol 14α-methyl
albicans enzyme has been proposed (24). The crystal structure demethylase cytochrome P450.
of the enzyme from Mycobacterium tuberculosis was recently
published (PDB ID 1E9X and 1EA1) (25). This is the first
enzyme in this pathway for which there is a crystal structure.
The enzyme is inhibited by compounds, such as
miconazole and ketoconazole (lipophilic imidazoles) (Fig. 3),
that coordinate through the imidazole nitrogen to the heme
iron of the cytochrome P450 and block binding of O2 (14,
25). The result is an accumulation of dihydrolanosterol (the
C24 reduction product of lanosterol).
Reduction of 5 to 6 HO 5
H* Hw
The reduction of cholestatriene (5) to 14-demethyl- HO
6
lanosterol (4,4′-dimethylzymosterol; 4,4′-dimethylcholesta-
Hw+
8,24-dien-3β-ol) (6) is catalyzed by ∆14-sterol reductase (sterol
∆14-reductase; steroid-14-reductase; ∆8,14-sterol ∆14-reductase) NADPH*

(26, 27 ). The enzyme requires NADPH and the mechanism

involves formation of a carbocation intermediate (Fig. 4). A
proton from the aqueous solution (or from a general acid in
the enzyme active site) protonates at C15 to form a carbo- + Hw
cation, which is stabilized by the active site of the enzyme HO
and by the adjacent double bond. The enzyme directs
transfer of a hydride from NADPH to C14 that completes
the reduction reaction. Figure 4. Reduction of the C14 double bond by ∆14-sterol reductase.

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Oxidation of 6 to 9
The oxidation of 4,4′-dimethylzymosterol (6) to 4-
methylzymosterol-4-carboxylic acid (4β-methyl-4α-carboxy- NAD+
H
cholesta-8,24-dien-3β-ol) (9) is catalyzed by sterol-4α-methyl-
oxidase (C4 sterol methyl oxidase; C4 methyl sterol oxidase; O 9
H C O
4-methyl sterol oxidase; C4 methyl oxidase; C4-methyl- –
O
oxidase) (28, 29). The enzyme is specific for oxidation of the
C4α methyl group. It requires three NADH coenzymes and HO
three O2 molecules. The reducing equivalents from NADH are
acquired by NADH:cytochrome b5 reductase and cytochrome
b5. The enzyme catalyzes three successive (monooxygenation) CO2
oxidation reactions: (i) 4,4′-dimethylzymosterol (6) to 4-methyl-
4-hydroxymethylzymosterol (4β-methyl-4α-hydroxymethyl-
cholesta-8,24-dien-3β-ol) (7), (ii) 4-methyl-4-hydroxymethyl-
zymosterol (7) to 4-methylzymosterol-4-carboxaldehyde (4β- O
C O
methyl-4α-formyl-cholesta-8,24-dien-3β-ol) (8), and (iii) H O
O 10
4-methylzymosterol-4-carboxaldehyde (8) to 4-methyl- H
zymosterol-4-carboxylic acid (9). The human cDNA clone has
been mapped to chromosome 4q32-34, and histidine-rich Figure 5. Oxidative decarboxylation and enolization epimerization
clusters are suggested to be involved in the binding of oxo- of the 3β-hydroxycarboxylic acid by C3 sterol dehydrogenase (C4
diiron (Fe–O–Fe) (29). decarboxylase).

Oxidative Decarboxylation and Enolization

Epimerization of 9 to 10
The oxidative decarboxylation and enolization epimeriza-
tion of 4-methylzymosterol-4-carboxylic acid (9) to 4-methyl-
zymosterone (3-keto-4-methylzymosterol) (10) is catalyzed by
C3 sterol dehydrogenase (C4 decarboxylase) (4α-oic acid de-
carboxylase; C4 decarboxylase) (15). The enzyme requires an
NAD+ coenzyme. The enzyme catalyzes oxidation of 9 (Fig. 5) HO H 16
by transfer of the 3α hydrogen as a hydride to NAD+, leading H*
to a 3-ketocarboxylic acid intermediate that undergoes Hw+
decarboxylation to give an enol. The enzyme directs the epimer-
ization from the enol that moves the former 4β-methyl group
to the 4α-methyl position.
Hw
Reduction of 10 to 11 +
The reduction of 4-methylzymosterone (10) to 4α- HO H
methylzymosterol (4α-methyl-cholesta-8,24-dien-3β-ol) (11) is H* H*+
catalyzed by 3-keto reductase (3-ketosterol reductase; steroid-
3-ketoreductase) (16 ). The enzyme, which requires an HO 17
NAD(P)H coenzyme, catalyzes the 3-ketoreduction to convert
the 3-keto group to the β-hydroxy sterol.
Figure 6. The isomerization reaction catalyzed by ∆ 8-∆7 sterol
Oxidation of 11 to 14 isomerase.
The oxidation of 4 α -methylzymosterol (11) to
zymosterol-4α-carboxylic acid (4α-carboxy-cholesta-8,24-
dien-3β-ol) (14) is catalyzed by sterol-4α-methyl-oxidase, the
same enzyme that catalyzes the oxidation of 6 to 9. It cata-
lyzes three successive (monooxygenation) oxidation reactions: that catalyzes the oxidative decarboxylation and enolization
(i) 4 α -methylzymosterol (11) to 4 α -hydroxymethyl- epimerization of 9 to 10. The enzyme catalyzes oxidation of 14
zymosterol (4α -hydroxymethyl-cholesta-8,24-dien-3β-ol) to a 3-ketocarboxylic acid intermediate that undergoes decarbox-
(12), (ii) 4α-hydroxymethylzymosterol (12) to zymosterol- ylation to give an enol, which undergoes tautomerization.
4α-carboxaldehyde (4α-formyl-cholesta-8,24-dien-3β-ol)
(13), and (iii) zymosterol-4 α -carboxaldehyde (13) to Reduction of 15 to 16
zymosterol-4α-carboxylic acid (14). The reduction of zymosterone (15) to zymosterol (5α-
cholesta-8,24-dien-3 β -ol; 5 α -cholest-8-en-3 β -ol; ∆ 8,24-
Oxidative Decarboxylation of 14 to 15 cholestadien-3β-ol; ∆8-cholestenol) (16) is catalyzed by 3-keto
The oxidative decarboxylation of zymosterol-4α-carboxylic reductase, the same enzyme that catalyzes the reduction of
acid (14) to zymosterone (15) is catalyzed by C3 sterol de- 10 to 11. The enzyme catalyzes the 3-ketoreduction to convert
hydrogenase (C4 decarboxylase), which is the same enzyme the 3-keto group to the β-hydroxy sterol.

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 Research: Science and Education

Isomerization of 16 to 17
The isomerization of zymosterol (16) to lanthosterol (5α-
cholesta-7,24-dien-3 β -ol; 5 α -cholest-7-en-3 β -ol; ∆ 7,24-
HO
cholestadien-3β-ol; cholest-7-en-3β-ol) (17) is catalyzed by ∆8-∆7
Lanthosterol sterol isomerase (sterol ∆8-isomerase; 3β-hydroxysteroid-∆8-
HO ∆7-isomerase; cholestenol ∆-isomerase; ∆7-cholestenol ∆7-∆8-
NADH
O2
Desmosterol isomerase, EC 5.3.3.5) (30–33). The enzyme is also an
NADPH
emopamil-binding protein and a sigma factor. The human
cDNA clone has been reported (30). The enzyme catalyzes
NAD(P)H
isomerization of the double bond (Fig. 6) by protonation at C9
by a proton from the aqueous solution (or from a general acid
in the enzyme active site) to form a carbocation intermediate,
which is stabilized by the active site of the enzyme. The enzyme
HO directs subsequent abstraction of the 7β H to give the product.
7-Dehydrodesmosterol
This isomerization is the only reversible reaction in the 19
steps of biosynthesis of cholesterol from lanosterol (33).
HO
Cholesterol
Figure 7. The position in the pathway of C24 reduction has most Reduction of 17 to 18
often been placed as the last step in the conversion of lanos-
terol to cholesterol. In this position, the pathway from lanthosterol
The reduction of lanthosterol (17) to lathosterol (5α-
to cholesterol is shown. cholest-7-en-3β-ol; ∆7-cholesten-3β-ol) (18) is catalyzed by sterol
∆24-reductase (24-reductase; 3β-hydroxysterol ∆24-reductase)
(34). The exact location in the pathway for the C24 reduction
is not known because the enzyme catalyzes the reduction of the
double bond at each step in the pathway (e.g., see conversion
H* of 1 to 5). The position of the C24 reduction has most often
18
HO been placed as the last step in the pathway so that lanthosterol
H H is converted first to 7-dehydrodesmosterol and then to
desmosterol before final reduction of C24 to cholesterol (Fig.
H
HO
7). Bae and Paik (34) reported that the relative rates of re-
H H duction by sterol ∆24-reductase were 1.0 for lanthosterol (17),
H* 0.34 for zymosterol (16), 0.31 for desmosterol, and 0.06 for
lanosterol (1); lanthosterol is more easily reduced than
desmosterol or zymosterol. On the basis of their proposal from
this study, I have placed the C24 reduction at this location
in the pathway. The enzyme requires an NAD(P)H coenzyme
and presumably catalyzes the reduction by a carbocation
19
HO intermediate.
*
H
Figure 8. Lathosterol oxidase catalyzes the formation of the C5 Dehydrogenation (Desaturation) of 18 to 19
double bond.
The dehydrogenation (desaturation) of lathosterol (18) to
7-dehydrocholesterol (cholesta-5,7-dien-3β-ol) (19) is catalyzed
by lathosterol oxidase (lathosterol 5-desaturase; ∆7-sterol ∆5-
reductase; ∆7-sterol 5-reductase; ∆7-sterol-C5(6)-desaturase;
sterol 5α,6α-desaturase; sterol ∆5 desaturase; ∆5-dehydrogenase;
19
Hw C5-sterol desaturase; 5α-cholest-7-en-3β-ol:O2 ∆5-oxido-
HO reductase, EC 1.3.3.2) (35, 36 ). The enzyme requires an
20
HO
NAD(P)H coenzyme (NADH is better than NADPH) and
Hw+ H*
an O2 molecule; the reducing equivalents from NADH are
acquired by NADH:cytochrome b5 reductase and cytochrome
NADPH* b5. The enzyme contains histidine-rich motifs that would
provide the ligands for a presumed catalytic Fe center (35).
It catalyzes the stereospecific dehydrogenation (desaturation)
Hw of the 5α and 6α hydrogen atoms in a cis abstraction. A study
indicated an asynchronous scission of the two C–H bonds
+ (Fig. 8) where the C6α H is removed first to form a radical
HO
intermediate, and then the C5α H is removed, rather than a
Figure 9. The last step in the pathway is reduction of the C7 double concerted desaturation mechanism (36 ). The human cDNA
bond by 7-dehydrocholesterol reductase. clone has been reported (37 ).

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 Research: Science and Education
Reduction of 19 to 20
The reduction of 7-dehydrocholesterol (19) to cholesterol
(20) is catalyzed by 7-dehydrocholesterol reductase (∆7-sterol
reductase; 3β-hydroxysterol ∆7-reductase; 7-dehydrocholesterol
∆7-reductase; cholesterol:NADP + ∆7-oxidoreductase, EC
1.3.1.21) (38, 39). The enzyme requires an NADPH coenzyme.
It catalyzes ∆7-reduction (Fig. 9) by protonation at C8β by a
proton from aqueous solution (or from a general acid in the
enzyme active site) to form a carbocation intermediate that is
stabilized by the active site of the enzyme and by the adjacent
double bond. The enzyme directs hydride transfer from NADPH
to C7α that completes the reduction reaction. The human
cDNA clone has been reported (38) and mapped to chromo-
some 11q12-13 (40).
The 19-step conversion of lanosterol to cholesterol requires
nine different enzymes. It has been proposed to occur in both
the endoplasmic reticulum and the peroxisome compartments
of the cell (41). trans-1,4-Bis-(2-chlorobenzylamino-
methyl)cyclohexane dihydrochloride (AY-9944) is an inhibitor
of ∆14-sterol reductase (conversion of 5 to 6), ∆8–∆7 sterol
isomerase (conversion of 16 to 17), and 7-dehydrocholesterol
reductase (conversion of 19 to 20) with application in hyper-
lipidemia. trans-2-[4-(1,2-Diphenylbuten-1-yl)phenoxy]-
N,N-dimethylethylamine (tamoxifin), a chemotherapeutic
agent used in the treatment of breast cancer, is an inhibitor
of ∆8–∆7 sterol isomerase.
Cholesterol Biosynthesis in Perspective
The biosynthesis of cholesterol from lanosterol is not un-
like other metabolic pathways in that the enzymes catalyze a
series of oxidation and reduction reactions along with carbon–
carbon bond cleavage, epimerization, isomerization, and de-
hydrogenation. The five reduction reactions (5 → 6, 10 → 11,
15 → 16, 17 → 18, 19 → 20) utilize reduced flavoenzymes
(NAD[P]H) that are common in metabolic pathways (42). The
epimerization (part of 9 → 10) and isomerization (16 → 17)
reactions are also observed in other metabolic pathways (42).
Removal of the three methyl groups from lanosterol
(C30, C31, C32) requires carbon–carbon bond cleavage. To
accomplish this, the methyl groups are oxidized to carboxylic
acids and eliminated as a formic acid (4 → 5) and two carbon
dioxides (part of 9 → 10, 14 → 15). Three enzymes catalyze
the demethylation reactions. A cytochrome P450 enzyme is used
for one complete oxidative decarboxylation of C32 (1 → 5),
and two enzymes are used in the oxidation (6 → 9, 11 → 14)
and oxidative decarboxylation (9 → 10, 14 → 15) reactions
of C30 and C31.
Oxidations catalyzed by cytochrome P450 enzymes are
common (43–45), but oxidations ending in carbon–carbon
bond cleavage appear unique to certain steroid-metabolizing
cytochrome P450 enzymes (43, 44), including lanosterol
14α-methyl demethylase cytochrome P450 here. Thus, this
type of carbon–carbon bond cleavage is not widely seen in
metabolic pathways. On the other hand, the oxidation reactions
of C30 and C31 are thought to involve non-heme oxo-diiron
(29). The mechanism for oxidation by these binuclear non-
heme iron centers is believed to be formally equivalent to
that of cytochrome P450 (36, 42, 46 ), and enzymes utilizing
non-heme oxo-diiron centers are being more widely identified
(36, 46 ). But the reaction here does not end in carbon–carbon
382 Journal of Chemical Education • Vol. 79 No. 3 March 2002 • JChemEd.chem.wisc.edu
bond cleavage. Instead, a second enzyme catalyzes oxidation of
the β-hydroxyacid to a β-ketoacid, which undergoes decar-
boxylation. Decarboxylation via a β-ketoacid is more familiar
in metabolic pathways, as, for example, in the citric acid cycle
where isocitrate dehydrogenase catalyzes oxidative decarboxyla-
tion of isocitrate (via oxalosuccinate) to α-ketoglutarate, and
in the oxidative phase of the pentose phosphate pathway
where 6-phosphogluconate dehydrogenase catalyzes oxidative
decarboxylation of 6-phosphogluconate (via 6-phospho-3-oxo-
gluconate) to ribulose-5-phosphate. Therefore, the biosynthetic
pathway for cholesterol from lanosterol displays two signifi-
cantly different mechanisms for carbon–carbon bond cleavage;
the mechanisms are related via the oxidation reactions using
iron centers.
Finally, the dehydrogenation (desaturation) reaction (18 →
19) is not catalyzed by a flavoenzyme, although these are wide-
spread in metabolic pathways (e.g., succinate dehydrogenase
in the citric acid cycle, fatty acyl-CoA dehydrogenase in the
β-oxidation of fatty acids, and dihydroorotate oxidase in py-
rimidine nucleotide biosynthesis). Instead, a reduced flavin
(NAD[P]H) and O2 are involved, presumably with a non-
heme iron (oxo-diiron) center. These non-heme iron enzymes
participate in the biosynthesis of unsaturated fatty acids as
fatty acyl-CoA desaturases (e.g., ∆6-desaturase, ∆9-stearoyl-
CoA desaturase, ∆11-myristoyl-CoA desaturase, and ∆12-oleyl-
CoA desaturase) (36 ). Thus, this type of dehydrogenation
may be primarily associated with lipid metabolism.
Cholesterol Homeostasis
Much has been learned about cholesterol homeostasis,
although a complete picture has not yet emerged. Central to
cholesterol homeostasis are the sterol regulatory element-
binding proteins (SREBPs) (particularly SREBP-2, but also
SREBP-1c), which are under sterol regulation. SREBP-1a and
-1c primarily regulate fatty acid biosynthesis (47–49). When intra-
cellular cholesterol levels decline, SREBP-cleavage activating
protein (SCAP) forms a complex with SREBP that mediates the
interaction of SREBP with site-1-protease (S1P). A recent study
indicated that modifications of the N-linked carbohydrate
moieties on SCAP are important for this to occur (50).
S1P is a membrane-bound subtilisin-related serine protease
(51) that catalyzes the first proteolytic reaction on SREBP.
Site-2-protease (S2P), a member of the family of zinc metallo-
proteases (51), catalyzes hydrolysis of the second proteolytic
reaction on SREBP, which releases a soluble transcription factor
(the amino-terminal fragment of SREBP). The soluble tran-
scription factor enters the nucleus, where (usually with a second
transcription factor such as Sp-1 or NF-1) it binds to direct
repeat sterol response elements (SRE) in promotor regions,
activating transcription of sterol-regulated cholesterogenic
genes. Genes under this regulatory control include, at least,
LDL receptor, HMG-CoA synthase, HMG-CoA reductase,
farnesyl diphosphate synthase, squalene synthase, lanosterol
synthase, and lanosterol 14α-methyl demethylase cytochrome
P450. Activation of these genes restores intracellular choles-
terol levels.
When intracellular cholesterol levels build up, the activa-
tion of SREBPs by proteolysis is suppressed, which decreases
gene transcription for cholesterol biosynthesis. Cholesterol
itself, when added to cultured cells, only weakly suppresses

cholesterol biosynthesis. However, when 25-hydroxycholes-
terol (cholest-5-ene-3β,25-diol), an oxysterol (52) synthesized
by cholesterol 25-hydroxylase (53), is added to cultured cells
there is a rapid decrease in the activation of SREBPs and a
consequent rapid decrease in cholesterol biosynthesis. If 25-
hydroxycholesterol is the regulatory agent, the recent study
indicates that the sterol blocks modification of the N-linked
carbohydrate moieties of SCAP, which precludes an interaction
between S1P and SREBP (50).
Oxysterols (formed by hydroxylation of the side chain
of cholesterol) are modulators of numerous processes (52).
They are important in cholesterol homeostasis in peripheral
tissues such as brain and kidney where excess cholesterol is
converted to oxysterols that are more hydrophilic than cho-
lesterol. These are transported to the liver for conversion to
bile acids, which is under control of nuclear orphan receptors
(54). Bile acids are synthesized by two biosynthetic pathways:
from cholesterol (the “classical” pathway) and from oxysterols
(a newly described pathway) (54).
Inherited Diseases of Cholesterol Biosynthesis
Deficiencies in four enzymes of cholesterol biosynthesis
from lanosterol have been reported to give rise to diseases.
Mutations have been identified in the genes for three of these
enzymes. Mutations in C3 sterol dehydrogenase (C4 decarbox-
ylase) (conversion of 9 to 10, 14 to 15) give rise to CHILD
(congenital hemidysplasia, ichthyosis, and limb defects)
syndrome (55) (MIM 308050 [56 ]). CHILD syndrome is
characterized by dry, thickened, scaly skin lesions (“fishskin”)
on one side of the body; limb defects; calcification points on
cartilage structures; and internal organ anomalies.
Mutations in ∆8–∆7 sterol isomerase (conversion of 16
to 17) give rise to two forms of chondrodysplasia punctata
(CDP) (57–59). CDP describes calcification points on car-
tilage structures and disorders of bone growth and structure
associated with skeletal dwarfism. One form of CDP is
Conradi–Hünermann syndrome (CDPX2) (MIM 302960
[56 ]), which arises by mutations in ∆8–∆7 sterol isomerase (58).
It resembles CHILD syndrome, but the dry, thickened, scaly
skin lesions are on both sides of the body in an asymmetric
pattern. Because CHILD syndrome and CDPX2 have similar
clinical features, a patient diagnosed with CHILD syndrome was
reported to have a mutation in the ∆8–∆7 sterol isomerase gene
(59). Thus CHILD syndrome and CDPX2 were proposed
to result from allelic mutations in the ∆8–∆7 sterol isomerase
gene. However, because CHILD syndrome was shown to arise
by mutations in C3 sterol dehydrogenase (C4 decarboxylase)
(55), the mutation in ∆8–∆7 sterol isomerase (59) may be
another form of CDP. Therefore, CHILD syndrome is distinct
from Conradi–Hünermann (with unilateral skin lesions and
bilateral skin lesions).
A deficiency in sterol ∆24-reductase (conversion of 17
to 18) activity leading to an accumulation of desmosterol has
been proposed to give rise to desmosterolosis (60). This disease
is characterized by abnormally large cranial capacity, cleft
palate, ambiguous genitalia, and short limbs.
Mutations of 7-dehydrocholesterol reductase (conversion
of 19 to 20) give rise to Smith–Lemli–Opitz syndrome (SLOS)
(40, 61–63) (MIM 270400 [56 ]). SLOS is characterized by
abnormal smallness of the head, poor growth, cleft palate,
JChemEd.chem.wisc.edu • Vol. 79 No. 3 March 2002 • Journal of Chemical Education
Research: Science and Education
malformation of the face, limb abnormalities, ambiguous
genitalia, and mental retardation. There are two phenotypes:
type I (mild) and type II (severe). The incidence of this auto-
somal recessive disorder is estimated to be 1 in 20,000 births.
It may be the second most common autosomal recessive dis-
order in the white population of North Americans (after cystic
fibrosis). Approximately three-quarters of an issue of the
American Journal of Medical Genetics was recently devoted to
SLOS (64 ). Recognition of cholesterol’s essential role in
mammalian embryonic development (1) established SLOS
as the prototypical developmental disorder (62).
Acknowledgments
I wish to thank James C. Crosthwaite, Thomas W.
Mattingly Jr., and Joanna K. Krueger for a critical reading of
the manuscript.
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