Document Detail


A mammalian mutant cell lacking detectable lanosterol 14 alpha-methyl demethylase activity.
MedLine Citation:
PMID:  3335544     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A Chinese hamster ovary cell mutant, AR45, was selected for amphotericin B resistance after treatment with the mutagen ethyl methanesulfonate. The mutant is a cholesterol auxotroph with a deficiency in cholesterol biosynthesis. Whole cell experiments demonstrate that the mutant accumulates the C30 sterols, lanosterol and dihydrolanosterol, under culture conditions which promote active sterol biosynthesis. Metabolic studies show that the C29 sterol demethylation product of lanosterol, but not lanosterol itself, is actively converted to end product cholesterol by whole cells as well as by microsomal preparations derived from the mutant. Detectable amounts of several cytochromes can be observed spectrally in the AR45 demonstrating that it is not a general heme-deficient mutant. Collectively, these results characterize the AR45 mutant cells as being lanosterol 14 alpha-methyl demethylase-deficient. The cell line should prove useful in studying regulation of the demethylase enzyme and the putative endogenous regulatory oxysterol. It should also be a useful tool in the molecular cloning and elucidation of genetic properties of the demethylase.
Authors:
H W Chen; D A Leonard; R T Fischer; J M Trzaskos
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  263     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1988 Jan 
Date Detail:
Created Date:  1988-02-25     Completed Date:  1988-02-25     Revised Date:  2010-04-06    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1248-54     Citation Subset:  IM    
Affiliation:
Medical Products Department, E. I. du Pont de Nemours and Company, Wilmington, Delaware 19898.
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MeSH Terms
Descriptor/Qualifier:
Amphotericin B / pharmacology
Animals
Cell Line
Cholesterol / analysis
Chromatography, Thin Layer
Cricetinae
Cytochrome P-450 Enzyme System*
Drug Resistance
Ethyl Methanesulfonate / pharmacology
Gas Chromatography-Mass Spectrometry
Hydroxymethylglutaryl CoA Reductases / metabolism
Mutation
Oxidoreductases / analysis*,  genetics
Phospholipids / analysis
Chemical
Reg. No./Substance:
0/Phospholipids; 1397-89-3/Amphotericin B; 57-88-5/Cholesterol; 62-50-0/Ethyl Methanesulfonate; 9035-51-2/Cytochrome P-450 Enzyme System; EC 1.-/Oxidoreductases; EC 1.1.1.-/Hydroxymethylglutaryl CoA Reductases; EC 1.14.13.70/sterol 14-demethylase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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