The Escherichia coli outer membrane protein OmpA acquires secondary structure prior to its integration into the membrane.
The Escherichia coli outer membrane protein OmpA acquires secondary structure prior to its integration into the membrane.
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DOI：
10.1016/j.jbc.2022.101802
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发表时间：
2022-04
期刊：
影响因子：
--
通讯作者：
Bernstein HD
中科院分区：
文献类型：
--
作者：
Wang X;Bernstein HD
几乎所有存在于革兰氏阴性细菌外膜 (OM) 中的蛋白质都含有跨膜片段，该片段折叠成独特的 β 桶结构，并通过未知机制插入膜中。为了进一步了解外膜蛋白 (OMP) 的生物发生，我们回顾了 20 多年前报道的令人惊讶的观察结果，即当大肠杆菌 OmpA β 桶表达为两个非共价连接的片段时，它可以在体内组装成天然结构。在这里，我们表明，N 端片段中的 β 链 4 和 C 端片段中的 β 链 5 之间的二硫键可以在周质空间中形成，并大大提高“分裂”OmpA 的组装效率，但前提是半胱氨酸残基以完美的方式进行设计（即，它们在完全折叠的 β 桶中对齐）。相比之下，我们观察到 N 端片段中的 β 链 1 和 C 端片段中的 β 链 8 之间仅存在微弱的二硫键，从而形成闭合或环状排列的 β 桶。我们的结果不仅证明 β 桶在整合到 OM 中之前开始折叠成 β-片状结构，而且还有助于区分已提出的不同的 OMP 生物发生模型。
Almost all proteins that reside in the outer membrane (OM) of Gram-negative bacteria contain a membrane-spanning segment that folds into a unique β barrel structure and inserts into the membrane by an unknown mechanism. To obtain further insight into outer membrane protein (OMP) biogenesis, we revisited the surprising observation reported over 20 years ago that the Escherichia coli OmpA β barrel can be assembled into a native structure in vivo when it is expressed as two noncovalently linked fragments. Here, we show that disulfide bonds between β strand 4 in the N-terminal fragment and β strand 5 in the C-terminal fragment can form in the periplasmic space and greatly increase the efficiency of assembly of “split” OmpA, but only if the cysteine residues are engineered in perfect register (i.e., they are aligned in the fully folded β barrel). In contrast, we observed only weak disulfide bonding between β strand 1 in the N-terminal fragment and β strand 8 in the C-terminal fragment that would form a closed or circularly permutated β barrel. Our results not only demonstrate that β barrels begin to fold into a β-sheet-like structure before they are integrated into the OM but also help to discriminate among the different models of OMP biogenesis that have been proposed.
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影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
DOI：
10.1126/science.aad3460
发表时间：
2016-01-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bakelar J;Buchanan SK;Noinaj N
通讯作者：
Noinaj N
影响因子：
56.9
作者：
Kim, Seokhee;Malinverni, Juliana C.;Kahne, Daniel
通讯作者：
Kahne, Daniel
DOI：
10.1073/pnas.1415955112
发表时间：
2015-02-17
影响因子：
11.1
作者：
Hagan, Christine L.;Wzorek, Joseph S.;Kahne, Daniel
通讯作者：
Kahne, Daniel