多种S1断裂内含肽间的体内交叉反应(英文)  

In Vivo Cross-reactivity between Multiple S1 Split Inteins

作　　者：宋慧玲[1,2] 刘相钦 孟清[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620 [2]戴尔蒙西大学生物化学与分子生物学系,Halifax,Nova Scotia B3H 4R2,Canada

出　　处：《中国生物化学与分子生物学报》2013年第8期734-739,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：Supported by National Science and Engineering Research Council grant of Canada;National High Technology Research and Development Program of China(863 Program,No.2006AA03Z451);National Natural Science Foundation of China(No.31070698);Shanghai Key Projects of Basic Research(No.10JC1400300);the Ph.D.Programs Foundation of Ministry of Education of China(No.20120075110007)~~

摘　　要：断裂内含肽含有两个独立分离的多肽片段(N端内含肽和C端内含肽),它催化蛋白质反式剪接反应,在蛋白质研究与蛋白质工程中已得到诸多实际应用.在蛋白质反式剪接过程中,内含肽的N端内含肽和C端内含肽通过结构互补特异性地非共价组合.然而,Ssp DnaX S1型断裂内含肽的较大C端内含肽片段近来被发现能够与源自其它内含肽的N端内含肽片段交叉反应,表明蛋白质内含子Ssp DnaX具有结构杂交特征.本研究对另外2种S1型内含肽RmaDnaB和Ssp Gyr B的较大C端内含肽与不同S1型断裂内含肽的N-端内含肽交叉反应活性进行分析检测.目的是探讨S1型断裂内含肽的结构杂交特征是否具有普遍性.结果发现,Rma DnaB的S1C端内含肽能够与Ssp GyrB的S1 N端内含肽交叉反应,却不能与Ssp DnaX的S1N端内含肽交叉反应;与此相似,Ssp GyrB的S1C端内含肽能够与Rma DnaB的S1 N端内含肽交叉反应,却不能与Ssp DnaX的S1N端内含肽交叉反应.此外,某些交叉反应表现出温度依赖性.这些结果对于内含肽的结构-功能关系以及S1型断裂内含肽的应用研究具有重要的意义.Split inteins consist of two separate polypeptides （N-intein and C-intein） and catalyze aprotein trans-splicing reaction, and have many practical applications in protein research and engineering. In protein trans-splicing, the N-intein and the C-intein assemble non-covalently and specifically through structural complementation. However, the large C-intein of Ssp DnaX S1 split intein was recently found to cross-react with putative N-inteins derived from other inteins, indicating a structural promiscuity of the Ssp DnaX intein. Here we evaluated two additional C-inteins, Rma DnaB and Ssp GyrB S1 split inteins, for cross-reactivity with N-inteins of different S1 split inteins, in order to determine whether S1 split inteins are generally promiscuous. We found that the C-intein of Rma DnaB S1 split intein could crossreact with the N-intein of Ssp GyrB S1 split intein, but not with the N-intein of Ssp DnaX S1 split intein. Similarly, the C-intein of Ssp GyrB S1 split intein could cross-react with the N-intein of Rma DnaB S1 split intein, but not with the N-intein of Ssp DnaX S1 split intein. Some of the cross-reactivities also showed a temperaturedependence. These findings have interesting implications on intein structurefunction and on applications of these S1 split inteins in developing intein-based technologies.

关 键 词：断裂内含肽 蛋白质反式剪接 交叉反应 

分 类 号：Q71[生物学—分子生物学]