Structure and regulation of mammalian squalene synthase.

Tansey TR1, Shechter I.

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Abstract

Mammalian squalene synthase (SQS) catalyzes the first reaction of the branch of the isoprenoid
metabolic pathway committed specifically to sterol biosynthesis. SQS produces squalene in an
unusual two-step reaction in which two molecules of farnesyl diphosphate are condensed head-
to-head. Recent studies have advanced understanding of the reaction mechanism, the functional
domains of the enzyme, and transcriptional regulation of the gene. Site-directed mutagenesis
has identified conserved Asp, Tyr, and Phe residues that are essential for SQS activity. The Asp
residues are hypothesized to be required for substrate binding; the Tyr and Phe residues may
stabilize carbocation reaction intermediates. The elucidation of SQS crystal structure will most
likely direct future research on the relationship between enzyme structure and function. SQS
activity, protein, and mRNA levels are regulated by cholesterol status and by the cytokines TNF-
alpha and IL-1beta. Activation of the SQS promoter in response to cholesterol deficit is mediated
by sterol regulatory element binding proteins SREBP-1a and SREBP-2. The precise contributions
made by individual SREBPs and accessory transcription factors to SQS transcriptional control,
and the mechanisms underlying cytokine regulation of SQS are major foci of current research.

Squalene synthase: structure and regulation.

Tansey TR1, Shechter I.

Author information

Abstract

Squalene synthase (SQS) catalyzes the first reaction of the branch of the isoprenoid metabolic
pathway committed specifically to sterol biosynthesis. Regulation of SQS is thought to direct
proximal intermediates in the pathway into either sterol or nonsterol branches in response to
changing cellular requirements. The importance of SQS in cholesterol metabolism has stimulated
research on the mechanism, structure, and regulation of the enzyme. SQS produces squalene, a
C30 isoprenoid, in a two-step reaction in which two molecules of farnesyl diphosphate are
condensed head to head. Site-directed mutagenesis of rat SQS has identified conserved Tyr, Phe,
and Asp residues that are essential for function. The aromatic rings of Tyr and Phe are postulated
to stabilize carbocation intermediates of the first and second half-reactions, respectively; the
acidic Asp residues may be required for substrate binding. SQS activity, protein level, and gene
transcription are strictly and coordinately regulated by cholesterol status, decreasing with
cholesterol surfeit and increasing with cholesterol deficit. The human SQS (hSQS) gene has an
unusually complex promoter with multiple binding sites for the sterol regulatory element
binding proteins SREBP-1a and SREBP-2, and for accessory transcription factors known to be
involved in the control of other sterol-responsive genes. SREBP-1a and SREBP-2 require different
subsets of hSQS regulatory DNA elements to achieve maximal promoter activation. Current
research is directed at elucidating the precise contribution made by individual SREBPs and
accessory transcription factors to hSQS transcriptional control.

Crystal structure of human squalene synthase. A key enzyme in cholesterol biosynthesis.

Pandit J1, Danley DE, Schulte GK, Mazzalupo S, Pauly TA, Hayward CM, Hamanaka ES, Thompson
JF, Harwood HJ Jr.

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Abstract

Squalene synthase catalyzes the biosynthesis of squalene, a key cholesterol precursor, through a
reductive dimerization of two farnesyl diphosphate (FPP) molecules. The reaction is unique
when compared with those of other FPP-utilizing enzymes and proceeds in two distinct steps,
both of which involve the formation of carbocationic reaction intermediates. Because FPP is
located at the final branch point in the isoprenoid biosynthesis pathway, its conversion to
squalene through the action of squalene synthase represents the first committed step in the
formation of cholesterol, making it an attractive target for therapeutic intervention. We have
determined, for the first time, the crystal structures of recombinant human squalene synthase
complexed with several different inhibitors. The structure shows that SQS is folded as a single
domain, with a large channel in the middle of one face. The active sites of the two half-reactions
catalyzed by the enzyme are located in the central channel, which is lined on both sides by
conserved aspartate and arginine residues, which are known from mutagenesis experiments to
be involved in FPP binding. One end of this channel is exposed to solvent, whereas the other end
leads to a completely enclosed pocket surrounded by conserved hydrophobic residues. These
observations, along with mutagenesis data identifying residues that affect substrate binding and
activity, suggest that two molecules of FPP bind at one end of the channel, where the active
center of the first half-reaction is located, and then the stable reaction intermediate moves into
the deep pocket, where it is sequestered from solvent and the second half-reaction occurs. Five
alpha helices surrounding the active center are structurally homologous to the active core in the
three other isoprenoid biosynthetic enzymes whose crystal structures are known, even though
there is no detectable sequence homology.