X-ray structures of Clostridium perfringens sortase C with C-terminal cell wall sorting motif of LPST demonstrate role of subsite for substrate-binding and structural variations of catalytic site
X-ray structures of Clostridium perfringens sortase C with C-terminal cell wall sorting motif of LPST demonstrate role of subsite for substrate-binding and structural variations of catalytic site
复制标题
具有 LPST C 末端细胞壁分选基序的产气荚膜梭菌分选酶 C 的 X 射线结构证明了亚位点对底物结合和催化位点结构变化的作用
DOI：
10.1016/j.bbrc.2021.03.106
复制
发表时间：
2021
影响因子：
3.1
通讯作者：
Kamitori Shigehiro
中科院分区：
文献类型：
--
作者：
Tamai Eiji;Sekiya Hiroshi;Nariya Hirofumi;Katayama Seiichi;Kamitori Shigehiro
革兰氏阳性菌的皮利（Pili）是一种与细菌细胞壁共价结合的棒状蛋白，在细菌细胞与宿主组织的初始粘附和细菌定植中起重要作用。皮利是由C类分选酶（SrtC）（半胱氨酸转肽酶家族）催化的主要菌毛蛋白和次要菌毛蛋白聚合而形成的。革兰氏阳性菌产气荚膜梭菌（Clostridium perfringens）具有主要菌毛蛋白（CppA）、次要菌毛蛋白（CppB）和SrtC（CpSrtC）。CpSrtC识别具有5个氨基酸残基的C-末端细胞壁分选信号基序，CppA的LPSTG和CppB的LPETG，用于皮利的聚合。在这里，我们报告的生化分析，以检测体内的形成ofClostridium perfringenspili，和一种新的分子间的CpSrtC的底物-酶复合物的X-射线结构与序列的LPST在C-末端网站。结果表明，CpSrtC有一个底物结合的辅助聚合的皮利，和催化位点的结构变化，提供洞察酶的催化反应机制和亲和力的C-末端细胞壁分选信号基序序列。
Pili of Gram-positive bacteria are flexible rod proteins covalently attached to the bacterial cell wall, that play important roles in the initial adhesion of bacterial cells to host tissues and bacterial colonization. Pili are formed by the polymerization of major and minor pilins, catalyzed by class C sortase (SrtC), a family of cysteine transpeptidases. The Gram-positive bacteriumClostridium perfringenshas a major pilin (CppA), a minor pilin (CppB), and SrtC (CpSrtC). CpSrtC recognizes the C-terminal cell wall sorting signal motifs with five amino acid residues, LPSTG of CppA and LPETG of CppB, for the polymerization of pili. Here, we report biochemical analysis to detect the formation ofClostridium perfringenspiliin vivo, and the X-ray structure of a novel intermolecular substrate-enzyme complex of CpSrtC with a sequence of LPST at the C-terminal site. The results showed that CpSrtC has a subsite for substrate-binding to aid polymerization of pili, and that the catalytic site has structural variations, giving insights into the enzyme catalytic reaction mechanism and affinities for the C-terminal cell wall sorting signal motif sequences.