文章基本信息

标题：Molecular cloning and expression of rat liver bile acid CoA ligase
本地全文：下载
作者：Charles N. Falany ; Xiaowei Xie ; James B. Wheeler 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：2002
卷号：43
期号：12
页码：2062-2071
DOI：10.1194/jlr.M200260-JLR200
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：Bile acid CoA ligase (BAL) is responsible for catalyzing the first step in the conjugation of bile acids with amino acids. Sequencing of putative rat liver BAL cDNAs identified a cDNA (rBAL-1) possessing a 51 nucleotide 5′-untranslated region, an open reading frame of 2,070 bases encoding a 690 aa protein with a molecular mass of 75,960 Da, and a 138 nucleotide 3′-nontranslated region followed by a poly(A) tail. Identity of the cDNA was established by: 1 ) the rBAL-1 open reading frame encoded peptides obtained by chemical sequencing of the purified rBAL protein; 2 ) expressed rBAL-1 protein comigrated with purified rBAL during SDS-polyacrylamide gel electrophoresis; and 3 ) rBAL-1 expressed in insect Sf9 cells had enzymatic properties that were comparable to the enzyme isolated from rat liver. Evidence for a relationship between fatty acid and bile acid metabolism is suggested by specific inhibition of rBAL-1 by cis-unsaturated fatty acids and its high homology to a human very long chain fatty acid CoA ligase. In summary, these results indicate that the cDNA for rat liver BAL has been isolated and expression of the rBAL cDNA in insect Sf9 cells results in a catalytically active enzyme capable of utilizing several different bile acids as substrates.