Molecular cloning and expression of rat liver bile acid CoA ligase Published, JLR Papers in Press, September 16, 2002. DOI 10.1194/jlr.M200260-JLR200
Molecular cloning and expression of rat liver bile acid CoA ligase Published, JLR Papers in Press, September 16, 2002. DOI 10.1194/jlr.M200260-JLR200
复制标题
大鼠肝胆汁酸 CoA 连接酶的分子克隆和表达发表，JLR Papers in Press，2002 年 9 月 16 日。DOI 10.1194/jlr.M200260-JLR200
作者：
C. Falany;Xiao;J. Wheeler;Jin Wang;Michelle Smith;Dongning He;S. Barnes
胆汁酸辅酶A连接酶(BAL)负责催化胆汁酸与氨基酸结合的第一步。对推测的大鼠肝脏BAL cDNAs进行测序，发现了一个含有51个核苷酸的5‘-非翻译区、一个2,070个碱基的开放阅读框，编码一个690个氨基酸的蛋白质，其相对分子质量为75,960 Da，以及一个138个核苷酸的3’-非翻译区和一个Poly(A)尾巴。通过对纯化的rBAL蛋白进行化学测序获得rBAL-1开放阅读框编码的多肽，经SDS-聚丙烯酰胺凝胶电泳法检测rBAL-1蛋白的表达，以及在昆虫细胞Sf9中表达的rBAL-1具有与大鼠肝脏分离酶相似的酶性质。顺式不饱和脂肪酸对rBAL-1的特异性抑制及其与人类超长链脂肪酸辅酶A连接酶的高度同源性，表明了脂肪酸与胆汁酸代谢之间的关系。综上所述，这些结果表明，大鼠肝脏BAL的cDNA已被分离，并在昆虫Sf9细胞中表达rBAL，产生了一种能够利用几种不同胆汁酸作为底物的催化活性酶。
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.
登录
查看更多内容
DOI：
10.1016/s0952-3278(99)80007-6
发表时间：
1999-05
期刊：
Prostaglandins, leukotrienes, and essential fatty acids
影响因子：
--
作者：
P. Watkins;J. Pevsner;S. Steinberg
通讯作者：
P. Watkins;J. Pevsner;S. Steinberg
DOI：
10.1073/pnas.95.15.8625
发表时间：
1998-07-21
影响因子：
11.1
作者：
Hirsch, D;Stahl, A;Lodish, HF
通讯作者：
Lodish, HF
DOI：
10.1006/bbrc.1999.0510
发表时间：
1999-04-13
影响因子：
3.1
作者：
Steinberg, SJ;Wang, SJ;Watkins, PA
通讯作者：
Watkins, PA
影响因子：
2.9
作者：
BABBITT, PC;KENYON, GL;DUNAWAYMARIANO, D
通讯作者：
DUNAWAYMARIANO, D
人肝脏特异性超长链酰基辅酶 A 合成酶：第二种酶活性蛋白的 cDNA 克隆和表征。
DOI：
10.1006/mgme.1999.2883
发表时间：
1999
期刊：
Molecular genetics and metabolism.
影响因子：
--
作者：
Steinberg,SJ;Wang,SJ;McGuinness,MC;Watkins,PA
通讯作者：
Watkins,PA