酵母双杂交法筛选组织蛋白酶B相互作用蛋白质

Screening for Cathepsin B Interacting Proteins by Yeast Two-Hybrid System

组织蛋白酶B是肿瘤侵袭和转移密切相关的一种蛋白酶,研究其作用网络对于开发基于组织蛋白酶B的抗肿瘤药物非常重要。采用酵母双杂交技术构建相关文库,以人组织蛋白酶B作为诱饵蛋白质,寻找其在肿瘤细胞中的直接相互作用蛋白质。在84个初始阳性克隆中,共有15个能通过3个报告基因的检测。BLAST分析发现,在GenBank数据库中可以匹配7个基因,编码7种已知的蛋白质:BCL2-associated athanogene 6 (BAG6), filaminA,alpha(FLNA),ras homolog family member G (RHOG),metallothionein 2A (MT2A), ZXD family zinc finger C(ZXDC),DEP domain containing 7(DEPDC7),A kinase(PRKA) interacting protein 1(AKIP1)。

Cathepsin B is an enzymatic protein that was closely related with tumor cell invasion and metastasis.The network of Cathepsin B is very important for research and development of new anticancer drug,for example,new Cathepsin B inhibitor.Using yeast two hybrid technique,the proteins that interacting with Cathepsin B were identified.Results:After screening analysis,15 positive clones of putative interactions were obtained and they were identified by sequencing and utilizing of the data online(http://www.ncbi.nlm.nih.gov).There were 7 genes including BCL2-associated athanogene 6(BAG6),filamin A,alpha(FLNA), ras homolog family member G(RHOG), metallothionein 2A(MT2A), ZXD family zinc finger C(ZXDC), DEP domain containing 7(DEPDC7),A kinase(PRKA)interacting protein 1(AKIP1).