Photolabeling acyl CoA binding proteins in microsome preparations

FEBS Letters

Hare, James F¹ 

[1] Department of Biochemistry and Molecular Biology, Oregon Health Sciences University, Portland, Or. 97201, USA

关键词: Photolabeling;    Diacylglycerol acyltransferase;    Microsome;    Detergent;    ASACoA;    12-(5-iodo-4-azido-2-salicyl)amino]-dodecanoyl-coenzyme A;    DGAT;    diacylglycerol-acyltransferase;    MGAT;    monoaclyglycerol-acyltransferase;    SDS;    sodium dodecyl sulfate;    CoA;    coenzyme A;   

DOI  :  10.1016/0014-5793(95)01177-G

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To identify key enzymes that participate in acylglycerol metabolism, 12-[(5-iodo-4-azidosalicyl)amino]dodecanoylcoenzyme A (ASACoA) was emploued as a photoaffinity label for those enzymes that use fatty acyl CoA as a co-substrate. ASACoA inhibited diacylglycerol acyltransferase activity in liver microsomes and was incorporated into triacylglycerol in a microsome dependent reaction. When photoactivated, ASACoA labeled four proteins in rat liver (75, 54, 50 and 37 kDa) and in epididymal fat cell (75, 64, 54 and 37 kDa) microsomes. Photolabeling was sensitive to palmitoyl CoA. After solubilization in Triton X-114, all four proteins were concentrated into the detergent phase, indicating that they are integral membrane proteins.

【 授权许可】

Unknown