Substrate-bound and substrate-free outward-facing structures of a multidrug ABC exporter

被引：25

作者：

Chaptal, Vincent ^{[1
]}

Zampieri, Veronica ^{[1
,9
]}

Wiseman, Benjamin ^{[1
,2
,10
]}

Orelle, Cedric ^{[3
]}

Martin, Juliette ^{[4
]}

Nguyen, Kim-Anh ^{[5
]}

Gobet, Alexia ^{[1
]}

Di Cesare, Margot ^{[3
]}

Magnard, Sandrine ^{[1
]}

Javed, Waqas ^{[3
,8
]}

Eid, Jad ^{[1
]}

Kilburg, Arnaud ^{[1
]}

Peuchmaur, Marine ^{[6
]}

Marcoux, Julien ^{[7
]}

Monticelli, Luca ^{[4
]}

Hogbom, Martin ^{[2
]}

Schoehn, Guy ^{[8
]}

Jault, Jean-Michel ^{[3
]}

Boumendjel, Ahcene ^{[5
]}

Falson, Pierre ^{[1
]}

机构：

[1] Univ Lyon, Drug Resistance & Membrane Prot Grp, Mol Microbiol & Struct Biochem Lab, CNRS UMR 5086,IBCP, 7 Passage Vercors, F-69367 Lyon, France

[2] Stockholm Univ, Dept Biochem & Biophys, Arrhenius Labs Nat Sci, Stockholm, Sweden

[3] Univ Lyon, Bacterial Nucleotide Binding Prot Grp, Mol Microbiol & Struct Biochem Lab, CNRS UMR 5086,IBCP, 7 Passage Vercors, F-69367 Lyon, France

[4] Univ Lyon, Modeling Biol Macromol Grp, Mol Microbiol & Struct Biochem Lab, CNRS UMR 5086,IBCP, 7 Passage Vercors, F-69367 Lyon, France

[5] Univ Grenoble Alpes, LRB, INSERM, F-38000 Grenoble, France

[6] Univ Grenoble Alpes, CNRS, DPM UMR 5063, F-38041 Grenoble, France

[7] Univ Toulouse, Inst Pharmacol & Biol Struct IPBS, UPS, CNRS,UMR 5089, F-31000 Toulouse, France

[8] Univ Grenoble Alpes, IBS, CNRS, CEA, F-38000 Grenoble, France

[9] European Mol Biol Lab, 71 Ave Martyrs,CS 90181, F-38042 Grenoble 9, France

[10] Karolinska Inst, Dept Biosci & Nutr, Huddinge, Sweden

来源：

SCIENCE ADVANCES | 2022年 / 8卷 / 04期

基金：

瑞典研究理事会;

关键词：

BINDING CASSETTE TRANSPORTER; ATP-BINDING; P-GLYCOPROTEIN; BACILLUS-SUBTILIS; MOLECULAR-BASIS; RESISTANCE; FLEXIBILITY; DYNAMICS; MSBA; PRINCIPLES;

D O I：

10.1126/sciadv.abg9215

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：

Multidrug ABC transporters translocate drugs across membranes by a mechanism for which the molecular features of drug release are so far unknown. Here, we resolved three ATP-Mg2+-bound outward-facing conformations of the Bacillus subtilis (homodimeric) BmrA by x-ray crystallography and single-particle cryo-electron microscopy (EM) in detergent solution, one of them with rhodamine 6G (R6G), a substrate exported by BmrA when over-expressed in B. subtilis. Two R6G molecules bind to the drug-binding cavity at the level of the outer leaflet, between transmembrane (TM) helices 1-2 of one monomer and TM5'-6' of the other. They induce a rearrangement of TM1-2, highlighting a local flexibility that we confirmed by hydrogen/deuterium exchange and molecular dynamics simulations. In the absence of R6G, simulations show a fast postrelease occlusion of the cavity driven by hydrophobicity, while when present, R6G can move within the cavity, maintaining it open.

引用

页数：10

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