用[125I]辅酶A衍生物对仓鼠棕色脂肪组织线粒体进行光亲和标记：与解偶联蛋白和ADP/ATP载体的差异相互作用。

Photoaffinity labeling of hamster brown adipose tissue mitochondria by an [125I] coenzyme A derivative: differential interaction with the uncoupling protein and ADP/ATP carrier.

作者信息

Woldegiorgis G, Duff T, Contreras L, Shrago E, Ruoho A E

机构信息

Department of Medicine, University of Wisconsin, Madison 53706.

出版信息

Biochem Biophys Res Commun. 1989 Jun 15;161(2):502-7. doi: 10.1016/0006-291x(89)92627-2.

DOI:10.1016/0006-291x(89)92627-2

PMID:2544163

Abstract

We have recently synthesized an azido [125I] CoA photolabel, N-(3-iodo-4-azidophenyl propionamide) cysteinyl-5-(2'thiopyridyl cysteine) CoA that specifically labeled the ADP/ATP carrier in beef heart mitochondria. In this study brown adipose tissue mitochondria were photolabeled with the azido [125I] ACT-CoA derivative with or without inhibitors. SDS gel electrophoresis and autoradiography of the separated proteins revealed exclusive photolabeling of two polypeptides corresponding to the ADP/ATP carrier and uncoupling protein. In the presence of carboxyatracytloside only the 32 kD UCP was labeled by [125I] ACT-CoA, whereas preincubation with GDP resulted in exclusive photolabeling of the 30 kD ADP/ATP carrier. Palmitoyl CoA but not palmitic acid inhibited photolabeling of both polypeptides.

摘要

我们最近合成了一种叠氮基[¹²⁵I]辅酶A光标记物，即N-(3-碘-4-叠氮基苯基丙酰胺)半胱氨酰-5-(2'-硫代吡啶基半胱氨酸)辅酶A，它能特异性标记牛心线粒体中的ADP/ATP载体。在本研究中，棕色脂肪组织线粒体用叠氮基[¹²⁵I]辅酶A衍生物进行光标记，标记时添加或不添加抑制剂。对分离出的蛋白质进行SDS凝胶电泳和放射自显影显示，两种多肽被特异性光标记，它们分别对应于ADP/ATP载体和解偶联蛋白。在羧基阿曲库铵存在的情况下，只有32 kD的解偶联蛋白被[¹²⁵I]辅酶A标记，而预先用GDP孵育则导致30 kD的ADP/ATP载体被特异性光标记。棕榈酰辅酶A而非棕榈酸抑制了这两种多肽的光标记。