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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Activation of the Escherichia coli β-barrel assembly machine (Bam) is required for essential components to interact properly with substrate
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Activation of the Escherichia coli β-barrel assembly machine (Bam) is required for essential components to interact properly with substrate

机译:必须激活大肠杆菌β-桶装配机(Bam),以使必需成分与底物正确相互作用

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摘要

The outer membrane (OM) of Gram-negative bacteria such as Escherichia coli contains lipoproteins and integral β-barrel proteins (outer-membrane proteins, OMPs) assembled into an asymmetrical lipid bilayer. Insertion of β-barrel proteins into the OM is mediated by a protein complex that contains the OMP BamA and four associated lipoproteins (BamBCDE). The mechanism by which the Bam complex catalyzes the assembly of OMPs is not known. We report here the isolation and characterization of a temperature-sensitive lethal mutation, bamAE373K, which alters the fifth polypeptide transport-associated domain and disrupts the interaction between the BamAB and BamCDE subcomplexes. Suppressor mutations that map to codon 197 in bamD restore Bam complex function to wild-type levels. However, these suppressors do not restore the interaction between BamA and BamD; rather, they bypass the requirement for stable holocomplex formation by activating BamD. These results imply that BamA and BamD interact directly with OMP substrates.
机译:革兰氏阴性细菌(例如大肠杆菌)的外膜(OM)包含脂蛋白和组装成不对称脂质双层的完整β桶蛋白(外膜蛋白,OMP)。 β-桶状蛋白插入OM由包含OMP BamA和四个相关脂蛋白(BamBCDE)的蛋白复合物介导。 Bam络合物催化OMP组装的机制尚不清楚。我们在这里报告了对温度敏感的致死突变bamAE373K的分离和表征,该突变改变了第五个与多肽运输相关的结构域,并破坏了BamAB和BamCDE亚复合物之间的相互作用。映射至bamD密码子197的抑制子突变可将Bam复合体功能恢复至野生型水平。但是,这些抑制器不能恢复BamA与BamD之间的相互作用。相反,他们通过激活BamD绕过了稳定的完整络合物形成的要求。这些结果暗示BamA和BamD直接与OMP底物相互作用。

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