Purification of the Transmembrane Polypeptide Channel Complex of the Salmonella Flagellar Type III Secretion System
Purification of the Transmembrane Polypeptide Channel Complex of the Salmonella Flagellar Type III Secretion System
复制标题
沙门氏菌鞭毛III型分泌系统跨膜多肽通道复合体的纯化
DOI：
10.1007/978-1-0716-3060-0_1
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发表时间：
2023
影响因子：
--
通讯作者：
Minamino Tohru
中科院分区：
文献类型：
--
作者：
Kinoshita Miki;Namba Keiichi;Minamino Tohru
许多能动细菌利用鞭毛III型分泌系统（fT 3SS）在细胞表面构建鞭毛。fT 3SS由跨膜输出门复合物和细胞质ATP酶环复合物组成，所述跨膜输出门复合物充当质子/蛋白质反向转运体，其将质子流与鞭毛蛋白输出偶联，所述细胞质ATP酶环复合物充当输出门复合物的激活剂。三个跨膜蛋白，FliP，FliQ和FliR，形成出口门复合物的核心结构，并且该核心复合物作为多肽通道，允许鞭毛结构亚基跨细胞质膜易位。在这里，我们描述了过量生产，增溶和纯化的沙门氏菌FliP/FliQ/FliR复合物的方法。
Many motile bacteria employ the flagellar type III secretion system (fT3SS) to build the flagellum on the cell surface. The fT3SS consists of a transmembrane export gate complex, which acts as a proton/protein antiporter that couples proton flow with flagellar protein export, and a cytoplasmic ATPase ring complex, which works as an activator of the export gate complex. Three transmembrane proteins, FliP, FliQ, and FliR, form a core structure of the export gate complex, and this core complex serves as a polypeptide channel that allows flagellar structural subunits to be translocated across the cytoplasmic membrane. Here, we describe the methods for overproduction, solubilization, and purification of theSalmonellaFliP/FliQ/FliR complex.