Cook H A, Kumamoto C A
Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
J Bacteriol. 1999 May;181(10):3010-7. doi: 10.1128/JB.181.10.3010-3017.1999.
SecB is a cytosolic protein required for rapid and efficient export of particular periplasmic and outer membrane proteins in Escherichia coli. SecB promotes export by stabilizing newly synthesized precursor proteins in a nonnative conformation and by targeting the precursors to the inner membrane. Biochemical studies suggest that SecB facilitates precursor targeting by binding to the SecA protein, a component of the membrane-embedded translocation apparatus. To gain more insight into the functional interaction of SecB and SecA, in vivo, mutations in the secA locus that compensate for the export defect caused by the secB missense mutation secBL75Q were isolated. Two suppressors were isolated, both of which led to the overproduction of wild-type SecA protein. In vivo studies demonstrated that the SecBL75Q mutant protein releases precursor proteins at a lower rate than does wild-type SecB. Increasing the level of SecA protein in the cell was found to reverse this slow-release defect, indicating that overproduction of SecA stimulates the turnover of SecBL75Q-precursor complexes. These findings lend additional support to the proposed pathway for precursor targeting in which SecB promotes targeting to the translocation apparatus by binding to the SecA protein.
SecB是一种胞质蛋白,对于大肠杆菌中特定周质蛋白和外膜蛋白的快速高效输出是必需的。SecB通过将新合成的前体蛋白稳定在非天然构象并将前体靶向内膜来促进输出。生化研究表明,SecB通过与SecA蛋白(膜嵌入转运装置的一个组分)结合来促进前体靶向。为了更深入了解SecB和SecA的功能相互作用,在体内分离了secA基因座中可补偿由secB错义突变secBL75Q引起的输出缺陷的突变。分离到了两个抑制子,二者均导致野生型SecA蛋白过量产生。体内研究表明,SecBL75Q突变蛋白释放前体蛋白的速率低于野生型SecB。发现增加细胞中SecA蛋白的水平可逆转这种缓慢释放缺陷,表明SecA过量产生刺激了SecBL75Q-前体复合物的周转。这些发现为所提出的前体靶向途径提供了额外支持,即SecB通过与SecA蛋白结合促进靶向转运装置。