Hydrogen-deuterium exchange mass spectrometry captures distinct dynamics upon substrate and inhibitor binding to a transporter

Ruyu Jia, Chloe Martens (), Mrinal Shekhar, Shashank Pant, Grant A. Pellowe, Andy M. Lau, Heather E. Findlay, Nicola J. Harris, Emad Tajkhorshid, Paula J. Booth () and Argyris Politis ()
Additional contact information
Ruyu Jia: King’s College London
Chloe Martens: King’s College London
Mrinal Shekhar: University of Illinois at Urbana-Champaign
Shashank Pant: University of Illinois at Urbana-Champaign
Grant A. Pellowe: King’s College London
Andy M. Lau: King’s College London
Heather E. Findlay: King’s College London
Nicola J. Harris: King’s College London
Emad Tajkhorshid: University of Illinois at Urbana-Champaign
Paula J. Booth: King’s College London
Argyris Politis: King’s College London

Nature Communications, 2020, vol. 11, issue 1, 1-10

Abstract: Abstract Proton-coupled transporters use transmembrane proton gradients to power active transport of nutrients inside the cell. High-resolution structures often fail to capture the coupling between proton and ligand binding, and conformational changes associated with transport. We combine HDX-MS with mutagenesis and MD simulations to dissect the molecular mechanism of the prototypical transporter XylE. We show that protonation of a conserved aspartate triggers conformational transition from outward-facing to inward-facing state. This transition only occurs in the presence of substrate xylose, while the inhibitor glucose locks the transporter in the outward-facing state. MD simulations corroborate the experiments by showing that only the combination of protonation and xylose binding, and not glucose, sets up the transporter for conformational switch. Overall, we demonstrate the unique ability of HDX-MS to distinguish between the conformational dynamics of inhibitor and substrate binding, and show that a specific allosteric coupling between substrate binding and protonation is a key step to initiate transport.

Date: 2020
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41467-020-20032-3 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-20032-3

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-020-20032-3

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().