亚基12的分子克隆与表达：26S蛋白酶的一个非MCP且非ATP酶亚基

Molecular cloning and expression of subunit 12: a non-MCP and non-ATPase subunit of the 26 S protease.

作者信息

Dubiel W, Ferrell K, Dumdey R, Standera S, Prehn S, Rechsteiner M

机构信息

Institut für Biochemie, Universitätsklinikum Charité, Humboldt-Universität, Berlin, Germany.

出版信息

FEBS Lett. 1995 Apr 17;363(1-2):97-100. doi: 10.1016/0014-5793(95)00288-k.

DOI:10.1016/0014-5793(95)00288-k

PMID:7729561

Abstract

A cDNA encoding subunit 12 (S12) of human erythrocyte 26 S protease has been isolated, sequenced and expressed. The cDNA contains an open reading frame that encodes a 36.6 kDA protein 96% identical to mouse Mov-34 and 67% identical to its Drosophila melanogaster homolog. Based on the high degree of sequence identity between human S12, mouse and Drosophila Mov-34 proteins, we conclude that the Mov-34 gene product is a component of the 26 S protease. Antibodies produced against two S12 fragments, Met1-Tyr95 (S12f95) and Met1-Leu205 (S12f205), react with S12 transferred to nitrocellulose from SDS-PAGE. In contrast, after transfer from native gels, the epitope(s) recognized by anti-S12f205 is exposed in the regulatory complex but appears to be masked when the regulatory complex associates with the multicatalytic protease.

摘要

编码人红细胞26S蛋白酶亚基12（S12）的cDNA已被分离、测序和表达。该cDNA包含一个开放阅读框，其编码一种36.6kDa的蛋白质，与小鼠Mov-34的同源性为96%，与黑腹果蝇同源物的同源性为67%。基于人S12、小鼠和果蝇Mov-34蛋白之间高度的序列同一性，我们得出结论，Mov-34基因产物是26S蛋白酶的一个组成部分。针对两个S12片段，即Met1-Tyr95（S12f95）和Met1-Leu205（S12f205）产生的抗体，能与从SDS-PAGE转移至硝酸纤维素膜上的S12发生反应。相反，从天然凝胶转移后，抗S12f205识别的表位在调节复合物中暴露，但当调节复合物与多催化蛋白酶结合时，该表位似乎被掩盖。