Molecular Cloning and Expression of the Human Delta7-sterol Reductase

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1998 Mar 21

PMID 9465114

Citations 54

Authors

F F Moebius

B U Fitzky

J N Lee

Y K Paik

H Glossmann

Affiliations

Soon will be listed here.

Abstract

Inhibitors of the last steps of cholesterol biosynthesis such as AY9944 and BM15766 severely impair brain development. Their molecular target is the Delta7-sterol reductase (EC 1.3.1.21), suspected to be defective in the Smith-Lemli-Opitz syndrome, a frequent inborn disorder of sterol metabolism. Molecular cloning of the cDNA revealed that the human enzyme is a membrane-bound protein with a predicted molecular mass of 55 kDa and six to nine putative transmembrane segments. The protein is structurally related to plant and yeast sterol reductases. In adults the ubiquitously transcribed mRNA is most abundant in adrenal gland, liver, testis, and brain. The Delta7-sterol reductase is the ultimate enzyme of cholesterol biosynthesis in vertebrates and is absent from yeast. Microsomes from Saccharomyces cerevisiae strains heterologously expressing the human cDNA remove the C7-8 double bond in 7-dehydrocholesterol. The conversion to cholesterol depends on NADPH and is potently inhibited by AY9944 (IC50 0.013 microM), BM15766 (IC50 1.2 microM), and triparanol (IC50 14 microM). Our work paves the way to clarify whether a defect in the delta7-sterol reductase gene underlies the Smith-Lemli-Opitz syndrome.

Citing Articles

Cholesterol overload in macrophages drives metabolic dysfunction-associated steatohepatitis via inhibiting 7-dehydrocholesterol reductase in mice.

Li X, Wang K, Sun Y, Wang Y, Wu J, Dang Y J Transl Med. 2024; 22(1):1085.

PMID: 39614331 PMC: 11605946. DOI: 10.1186/s12967-024-05905-1.

Proteomics Analysis on the Effects of Oxidative Stress and Antioxidants on Proteins Involved in Sterol Transport and Metabolism in Human Telomerase Transcriptase-Overexpressing-Retinal Pigment Epithelium Cells.

Duncan R, Keightley A, Lopez A, Hall C, Koulen P Int J Mol Sci. 2024; 25(20).

PMID: 39456672 PMC: 11507349. DOI: 10.3390/ijms252010893.

Chemical Inhibition of Sterol Biosynthesis.

Peeples E, Mirnics K, Korade Z Biomolecules. 2024; 14(4).

PMID: 38672427 PMC: 11048061. DOI: 10.3390/biom14040410.

What can be lost? Genomic perspective on the lipid metabolism of Mucoromycota.

Sokolowska B, Orlowska M, Okrasinska A, Pilsyk S, Pawlowska J, Muszewska A IMA Fungus. 2023; 14(1):22.

PMID: 37932857 PMC: 10629195. DOI: 10.1186/s43008-023-00127-4.

Proteomics research of SARS-CoV-2 and COVID-19 disease.

Zhang N, Wang S, Wong C Med Rev (2021). 2023; 2(4):427-445.

PMID: 37724330 PMC: 10388787. DOI: 10.1515/mr-2022-0016.

References

Mercer E . Sterol biosynthesis inhibitors: their current status and modes of action. Lipids. 1991; 26(8):584-97. DOI: 10.1007/BF02536422. View

Worman H, Evans C, Blobel G . The lamin B receptor of the nuclear envelope inner membrane: a polytopic protein with eight potential transmembrane domains. J Cell Biol. 1990; 111(4):1535-42. PMC: 2116249. DOI: 10.1083/jcb.111.4.1535. View

Tint G, Irons M, Elias E, Batta A, Frieden R, Chen T . Defective cholesterol biosynthesis associated with the Smith-Lemli-Opitz syndrome. N Engl J Med. 1994; 330(2):107-13. DOI: 10.1056/NEJM199401133300205. View

Moebius F, Hanner M, Knaus H, Weber F, Striessnig J, Glossmann H . Purification and amino-terminal sequencing of the high affinity phenylalkylamine Ca2+ antagonist binding protein from guinea pig liver endoplasmic reticulum. J Biol Chem. 1994; 269(46):29314-20. View

Hanner M, Moebius F, Weber F, Grabner M, Striessnig J, Glossmann H . Phenylalkylamine Ca2+ antagonist binding protein. Molecular cloning, tissue distribution, and heterologous expression. J Biol Chem. 1995; 270(13):7551-7. DOI: 10.1074/jbc.270.13.7551. View

Alley T, Gray B, Lee S, Scherer S, Tsui L, Tint G . Identification of a yeast artificial chromosome clone spanning a translocation breakpoint at 7q32.1 in a Smith-Lemli-Opitz syndrome patient. Am J Hum Genet. 1995; 56(6):1411-6. PMC: 1801101. View

Batta A, Tint G, SHEFER S, Abuelo D, Salen G . Identification of 8-dehydrocholesterol (cholesta-5,8-dien-3 beta-ol) in patients with Smith-Lemli-Opitz syndrome. J Lipid Res. 1995; 36(4):705-13. View

SHEFER S, Salen G, Batta A, Honda A, Tint G, Irons M . Markedly inhibited 7-dehydrocholesterol-delta 7-reductase activity in liver microsomes from Smith-Lemli-Opitz homozygotes. J Clin Invest. 1995; 96(4):1779-85. PMC: 185814. DOI: 10.1172/JCI118223. View

Lecain E, Chenivesse X, Spagnoli R, Pompon D . Cloning by metabolic interference in yeast and enzymatic characterization of Arabidopsis thaliana sterol delta 7-reductase. J Biol Chem. 1996; 271(18):10866-73. DOI: 10.1074/jbc.271.18.10866. View

10.

Lennon G, Auffray C, Polymeropoulos M, Soares M . The I.M.A.G.E. Consortium: an integrated molecular analysis of genomes and their expression. Genomics. 1996; 33(1):151-2. DOI: 10.1006/geno.1996.0177. View

11.

Hanner M, Moebius F, Flandorfer A, Knaus H, Striessnig J, KEMPNER E . Purification, molecular cloning, and expression of the mammalian sigma1-binding site. Proc Natl Acad Sci U S A. 1996; 93(15):8072-7. PMC: 38877. DOI: 10.1073/pnas.93.15.8072. View

12.

Silve S, Dupuy P, Kaghad M, Chalon P, Rahier A, Taton M . Emopamil-binding protein, a mammalian protein that binds a series of structurally diverse neuroprotective agents, exhibits delta8-delta7 sterol isomerase activity in yeast. J Biol Chem. 1996; 271(37):22434-40. DOI: 10.1074/jbc.271.37.22434. View

13.

Wolf C, Chevy F, Pham J, Kolf-Clauw M, Citadelle D, Mulliez N . Changes in serum sterols of rats treated with 7-dehydrocholesterol-delta 7-reductase inhibitors: comparison to levels in humans with Smith-Lemli-Opitz syndrome. J Lipid Res. 1996; 37(6):1325-33. View

14.

Porter J, Young K, Beachy P . Cholesterol modification of hedgehog signaling proteins in animal development. Science. 1996; 274(5285):255-9. DOI: 10.1126/science.274.5285.255. View

15.

Chiang C, Litingtung Y, Lee E, Young K, Corden J, Westphal H . Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function. Nature. 1996; 383(6599):407-13. DOI: 10.1038/383407a0. View

16.

Schuler G, Boguski M, Stewart E, Stein L, Gyapay G, Rice K . A gene map of the human genome. Science. 1996; 274(5287):540-6. View

17.

Moebius F, Bermoser K, Reiter R, Hanner M, Glossmann H . Yeast sterol C8-C7 isomerase: identification and characterization of a high-affinity binding site for enzyme inhibitors. Biochemistry. 1996; 35(51):16871-8. DOI: 10.1021/bi961996m. View

18.

Kelley R . A new face for an old syndrome. Am J Med Genet. 1997; 68(3):251-6. DOI: 10.1002/(sici)1096-8628(19970131)68:3<251::aid-ajmg1>3.0.co;2-p. View

19.

Alley T, Scherer S, Huizenga J, Tsui L, Wallace M . Physical mapping of the chromosome 7 breakpoint region in an SLOS patient with t(7;20) (q32.1;q13.2). Am J Med Genet. 1997; 68(3):279-81. View

20.

Dehart D, Lanoue L, Tint G, Sulik K . Pathogenesis of malformations in a rodent model for Smith-Lemli-Opitz syndrome. Am J Med Genet. 1997; 68(3):328-37. DOI: 10.1002/(sici)1096-8628(19970131)68:3<328::aid-ajmg15>3.0.co;2-v. View