霍乱毒素对聚（ADP - 核糖）聚合酶的单（ADP - 核糖）基化作用。

Mono(ADP-ribosyl)ation of poly(ADP-ribose)polymerase by cholera toxin.

作者信息

Martinez M, Price S R, Moss J, Alvarez-Gonzalez R

机构信息

Department of Microbiology and Immunology, Texas College of Osteopathic Medicine/UNT, Fort Worth 76107-2690.

出版信息

Biochem Biophys Res Commun. 1991 Dec 31;181(3):1412-8. doi: 10.1016/0006-291x(91)92096-3.

DOI:10.1016/0006-291x(91)92096-3

PMID:1764092

Abstract

Poly(ADP-ribose)polymerase (PADPRP) was found to be an efficient protein acceptor for the arginine-specific ADP-ribosylation reaction catalyzed by cholera toxin (CT). The covalent modification of PADPRP was carried out with [32P]2'-dNAD as a selective mono(ADP-ribosyl)ation substrate. Mono(2'-dADP-ribosyl)ated-PADPRP was identified by autoradiographic analysis of the CT reaction products following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Addition of recombinant ADP-ribosylation factor (rARF), a small GTP-binding protein that stimulates the enzymatic activity of CT, enhanced the mono(2'-dADP-ribosyl)ation of PADPRP in a time- and substrate-dependent manner. In contrast, rARF did not change the ADP-ribose polymerizing activity of PADPRP. Peptide mapping mapping of [32P] labeled (2'-dADP-ribose)-PADPRP, following partial proteolysis with papain, revealed that the DNA-binding domain of PADPRP contained the mono(2'-dADP-ribosyl)ated arginine residue(s). Our results are consistent with the conclusion that PADPRP is susceptible to arginine-specific mono(ADP-ribosyl)ation catalyzed by CT.

摘要

聚（ADP - 核糖）聚合酶（PADPRP）被发现是霍乱毒素（CT）催化的精氨酸特异性ADP - 核糖基化反应的有效蛋白质受体。使用[32P]2'-dNAD作为选择性单（ADP - 核糖基）化底物对PADPRP进行共价修饰。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳后对CT反应产物的放射自显影分析鉴定单（2'-dADP - 核糖基）化的PADPRP。添加重组ADP - 核糖基化因子（rARF），一种刺激CT酶活性的小GTP结合蛋白，以时间和底物依赖性方式增强了PADPRP的单（2'-dADP - 核糖基）化。相反，rARF没有改变PADPRP的ADP - 核糖聚合活性。用木瓜蛋白酶进行部分蛋白水解后，对[32P]标记的（2'-dADP - 核糖） - PADPRP进行肽图谱分析，结果表明PADPRP的DNA结合结构域含有单（2'-dADP - 核糖基）化的精氨酸残基。我们的结果与PADPRP易受CT催化的精氨酸特异性单（ADP - 核糖基）化影响这一结论一致。