Hydrogen-deuterium exchange mass spectrometry captures distinct dynamics upon substrate and inhibitor binding to a transporter
| Autor: | Emad Tajkhorshid, Nicola J. Harris, Chloe Martens, Grant A. Pellowe, Heather E. Findlay, Andy M. Lau, Ruyu Jia, Paula J. Booth, Argyris Politis, Mrinal Shekhar, Shashank Pant |
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| Rok vydání: | 2020 |
| Předmět: |
Protein Conformation
alpha-Helical 0301 basic medicine Gene Expression General Physics and Astronomy 02 engineering and technology Crystallography X-Ray 01 natural sciences Substrate Specificity Membrane proteins Cloning Molecular 0303 health sciences Xylose Multidisciplinary Symporters 010304 chemical physics Chemistry Escherichia coli Proteins 021001 nanoscience & nanotechnology Recombinant Proteins Transmembrane protein 3. Good health Thermodynamics Permeation and transport Protons Structural biology 0210 nano-technology Sciences exactes et naturelles Protein Binding Science Genetic Vectors Allosteric regulation Hydrogen Deuterium Exchange-Mass Spectrometry Protonation Molecular Dynamics Simulation Article General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0103 physical sciences Escherichia coli Protein Interaction Domains and Motifs 030304 developmental biology Binding Sites Mass spectrometry Mutagenesis Glucose transporter Deuterium Exchange Measurement Substrate (chemistry) Transporter General Chemistry Kinetics Glucose 030104 developmental biology Mutation Biophysics Protein Conformation beta-Strand Hydrogen–deuterium exchange Xylose binding |
| Zdroj: | Jia, R, Martens, C, Shekhar, M, Pant, S, Pellowe, G, Lau, A M, Findlay, H, Harris, N, Tajkhorshid, E, Booth, P & Politis, A 2020, ' Hydrogen-deuterium exchange mass spectrometry captures distinct dynamics upon substrate and inhibitor binding to a transporter ', Nature Communications, vol. 11, 6162 . https://doi.org/10.1038/s41467-020-20032-3 Nature Communications Nature communications, 11 (1 Nature Communications, Vol 11, Iss 1, Pp 1-10 (2020) |
| Popis: | 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. SCOPUS: ar.j info:eu-repo/semantics/published |
| Databáze: | OpenAIRE |
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