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Volume 271, Number 37, Issue of September 13, 1996 pp. 22434-22440
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Emopamil-binding Protein, a Mammalian Protein That Binds a Series of Structurally Diverse Neuroprotective Agents, Exhibits Delta 8-Delta 7 Sterol Isomerase Activity in Yeast

(Received for publication, May 30, 1996)

Sandra Silve Dagger , Pascal Henry Dupuy Dagger , Christine Labit-Lebouteiller § , Mourad Kaghad , Pascale Chalon , Alain Rahier par , Maryse Taton par , Jan Lupker § , David Shire '' and Gérard Loison Dagger

From the Dagger  Department of Microbiology, the § Department of Animal Cell Technology, the  Department of Gene Molecular Biology, and the '' Department of Organic Chemistry, Sanofi-Recherche, Labège Innopole BP137, F-31676 Labège Cédex, France and the par  Department of Cellular and Molecular Enzymology, Institut de Botanique, 28 rue Goethe, F-67083, Strasbourg Cédex, France

Delta 8-Delta 7 sterol isomerase is an essential enzyme on the sterol biosynthesis pathway in eukaryotes. This endoplasmic reticulum-resident membrane protein catalyzes the conversion of Delta 8-sterols to their corresponding Delta 7-isomers. No sequence data for high eukaryote sterol isomerase being available so far, we have cloned a murine sterol isomerase-encoding cDNA by functional complementation of the corresponding deficiency in the yeast Saccharomyces cerevisiae. The amino acid sequence deduced from the cDNA open reading frame is highly similar to human emopamil-binding protein (EBP), a protein of unknown function that constitutes a molecular target for neuroprotective drugs. A yeast strain in which the sterol isomerase coding sequence has been replaced by that of human EBP or its murine homologue recovers the ability to convert Delta 8-sterol into Delta 7-sterol, both in vivo and in vitro. In these recombinant strains, both cell proliferation and the sterol isomerization reaction are inhibited by the high affinity EBP ligand trifluoperazine, as is the case in mammalian cells but not in wild type yeast cell. In contrast, the recombinant strains are much less susceptible to the sterol inhibition effect of haloperidol and fenpropimorph, as compared with wild type yeast strains. Our results strongly suggest that EBP and Delta 8-Delta 7 sterol isomerase are identical proteins in mammals.


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