The ABC transporter MsbA adopts the wide inward-open conformation in E. coli cells

被引：37

作者：

Galazzo, Laura ^{[1
,2
]}

Meier, Gianmarco ^{[3
]}

Januliene, Dovile ^{[4
]}

Parey, Kristian ^{[4
]}

De Vecchis, Dario ^{[5
]}

Striednig, Bianca ^{[3
]}

Hilbi, Hubert ^{[3
]}

Schaefer, Lars, V ^{[5
]}

Kuprov, Ilya ^{[6
]}

Moeller, Arne ^{[4
]}

Bordignon, Enrica ^{[1
,2
]}

Seeger, Markus A. ^{[3
]}

机构：

[1] Ruhr Univ Bochum, Fac Chem & Biochem, D-44801 Bochum, Germany

[2] Univ Geneva, Dept Phys Chem, CH-1211 Geneva, Switzerland

[3] Univ Zurich, Inst Med Microbiol, CH-8006 Zurich, Switzerland

[4] Osnabruck Univ, Dept Struct Biol, D-49076 Osnabruck, Germany

[5] Ruhr Univ Bochum, Ctr Theoret Chem, D-44801 Bochum, Germany

[6] Univ Southampton, Sch Chem, Southampton SO17 1BJ, Hants, England

来源：

SCIENCE ADVANCES | 2022年 / 8卷 / 41期

基金：

欧洲研究理事会; 瑞士国家科学基金会;

关键词：

BINDING CASSETTE TRANSPORTER; P-GLYCOPROTEIN REVEAL; ATP-BINDING; MOLECULAR-DYNAMICS; ESCHERICHIA-COLI; STRUCTURAL BASIS; CRYSTAL-STRUCTURE; CRYO-EM; FLEXIBILITY; NANOBODIES;

D O I：

10.1126/sciadv.abn6845

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Membrane proteins are currently investigated after detergent extraction from native cellular membranes and reconstitution into artificial liposomes or nanodiscs, thereby removing them from their physiological environment. However, to truly understand the biophysical properties of membrane proteins in a physiological environment, they must be investigated within living cells. Here, we used a spin-labeled nanobody to interrogate the conformational cycle of the ABC transporter MsbA by double electron-electron resonance. Unexpectedly, the wide inward-open conformation of MsbA, commonly considered a nonphysiological state, was found to be prominently populated in Escherichia coli cells. Molecular dynamics simulations revealed that extensive lateral portal opening is essential to provide access of its large natural substrate core lipid A to the binding cavity. Our work paves the way to investigate the conformational landscape of membrane proteins in cells.

引用

页数：15

共 21 条

[21] Comparative Analysis of Pulsed EPR Distance Measurements in an E. coli Cobalamin Transporter in Cells Versus Isolated Outer Membranes Reveals Novel Conformational Changes Dependent on the Native Environment
Nyenhuis, David
Nilaweera, Thushani
Cafiso, David S.
BIOPHYSICAL JOURNAL, 2019, 116 (03) : 128A - 128A

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