Journal Article

PUBDB-2020-03264

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Structural Kinetics of MsbA Investigated by Stopped-Flow Time-Resolved Small-Angle X-Ray Scattering

Josts, I.Extern*EMBL* ; Gao, Y. ; Monteiro, D. C. F. ; Niebling, S.EMBL* ; Nitsche, J. ; Veith, K.CSSB*EMBL* ; Gräwert, T. W.Extern*EMBL* ; Blanchet, C. E.Extern*EMBL* ; Schroer, M. A.Extern*EMBL* ; Huse, N. ; Pearson, A. R.Extern*EMBL* ; Svergun, D. I.EMBL* ; Tidow, H. (Corresponding author)Extern*EMBL*

2020
Cell Press Cambridge, Mass.

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Please use a persistent id in citations: doi:10.1016/j.str.2019.12.001  doi:10.3204/PUBDB-2020-03264

Abstract: Recent structures of full-length ATP-binding cassette (ABC) transporter MsbA in different states indicate large conformational changes during the reaction cycle that involve transient dimerization of its nucleotide-binding domains (NBDs). However, a detailed molecular understanding of the structural changes and associated kinetics of MsbA upon ATP binding and hydrolysis is still missing. Here, we employed time-resolved small-angle X-ray scattering, initiated by stopped-flow mixing, to investigate the kinetics and accompanying structural changes of NBD dimerization (upon ATP binding) and subsequent dissociation (upon ATP hydrolysis) in the context of isolated NBDs as well as full-length MsbA in lipid nanodiscs. Our data allowed us to structurally characterize the major states involved in the process and determine time constants for NBD dimerization and dissociation. In the full-length protein, these structural transitions occur on much faster time scales, indicating close-proximity effects and structural coupling of the transmembrane domains with the NBDs.

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Contributing Institute(s):

1. EMBL-User (EMBL-User)
2. CSSB-EMBL (CSSB-EMBL)
3. EMBL (EMBL)

Research Program(s):

1. 6G3 - PETRA III (POF3-622) (POF3-622)
2. DFG project 390715994 - EXC 2056: CUI: Advanced Imaging of Matter (390715994) (390715994)
3. DFG project 194651731 - EXC 1074: Hamburger Zentrum für ultraschnelle Beobachtung (CUI): Struktur, Dynamik und Kontrolle von Materie auf atomarer Skala (194651731) (194651731)

Experiment(s):

1. PETRA Beamline P12 (PETRA III)

Appears in the scientific report 2020

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Database coverage:
; ; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; Current Contents - Life Sciences ; Ebsco Academic Search ; Essential Science Indicators ; NCBI Molecular Biology Database ; Nationallizenz ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Web of Science Core Collection

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