Liberation of SARS-CoV main protease from the viral polyprotein:N-terminal autocleavage does not depend on the mature dimerization mode

作     者：Shuai Chen Felix Jonas Can Shen Rolf Higenfeld 

作者机构：Institute of BiochemistryCenter for Structural and Cell Biology in MedicineUniversity of LübeckRatzeburger Allee 16023538 LübeckGermany Laboratory for Structural Biology of Infection and Inflammationc/o DESYBuilding 22aNotkestr.8522603 HamburgGermany 

出 版 物：《Protein & Cell》 (蛋白质与细胞（英文版）)

年 卷 期：2010年第1卷第3期

页      面：59-74页

核心收录：

学科分类：0710[理学-生物学] 0831[工学-生物医学工程（可授工学、理学、医学学位）] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 0703[理学-化学] 0836[工学-生物工程] 10[医学] 

基　　金：This work was supported,in part,by the Sino-European Project on SARS Diagnostics and Antivirals(SEPSDA,contract NO.SP22-CT-2004-003831) by VIZIER(contract no.LSHG-CT-2004-511960) both funded by the European Commission.We acknowledge support from the Sino-German Center for the Promotion of Research,Beijing(grant no.233(202/6)) the Schleswig-Holstein Innovation Fund,and the DFG(Hi 611/4 and Cluster of Excellence“Inflammation at Interfaces”) 

主　　题：SARS-CoV M^(pro) N-terminal autocleavage autocleavage activity trans-cleavage activity dimerization 

摘      要：The main protease(M^(pro))plays a vital role in proteolytic processing of the polyproteins in the replicative cycle of SARS coronavirus(SARS-CoV).Dimerization of this enzyme has been shown to be indispensable for transcleavage ***,the auto-processing mechanism of M^(pro),*** own release from the polyproteins through autocleavage,remains *** study elucidates the relationship between the N-terminal autocleavage activity and the dimerization of“immatureM^(pro).Three residues(Arg4,Glu290,and Arg298),which contribute to the active dimer conformation of mature M^(pro),are selected for mutational ***,all three mutants still perform N-terminal autocleavage,while the dimerization of mature protease and transcleavage activity following auto-processing are completely inhibited by the E290R and R298E mutations and partially so by the R4E ***,the mature E290R mutant can resume N-terminal autocleavage activity when mixed with the“immatureC145A/E290R double mutant whereas its trans-cleavage activity remains ***,the N-terminal auto-processing of M^(pro) appears to require only two“immaturemonomers approaching one another to form an“intermediatedimer structure and does not strictly depend on the active dimer conformation existing in mature *** conclusion,an auto-release model of M^(pro) from the polyproteins is proposed,which will help understand the auto-processing mechanism and the difference between the autocleavage and trans-cleavage proteolytic activities of SARS-CoV M^(pro).