Simmons, KJ orcid.org/0000-0003-4846-9097, Jackson, SM, Brueckner, F et al. (14 more authors) (2014) Molecular mechanism of ligand recognition by membrane transport protein, Mhp1. The EMBO Journal, 33 (16). pp. 1831-1844. ISSN 0261-4189
Abstract
The hydantoin transporter Mhp1 is a sodium-coupled secondary active transport protein of the nucleobase-cation-symport family and a member of the widespread 5-helix inverted repeat superfamily of transporters. The structure of Mhp1 was previously solved in three different conformations providing insight into the molecular basis of the alternating access mechanism. Here, we elucidate detailed events of substrate binding, through a combination of crystallography, molecular dynamics, site-directed mutagenesis, biochemical/biophysical assays, and the design and synthesis of novel ligands. We show precisely where 5-substituted hydantoin substrates bind in an extended configuration at the interface of the bundle and hash domains. They are recognised through hydrogen bonds to the hydantoin moiety and the complementarity of the 5-substituent for a hydrophobic pocket in the protein. Furthermore, we describe a novel structure of an intermediate state of the protein with the external thin gate locked open by an inhibitor, 5-(2-naphthylmethyl)-L-hydantoin, which becomes a substrate when leucine 363 is changed to an alanine. We deduce the molecular events that underlie acquisition and transport of a ligand by Mhp1.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2014, The Authors. This is an Open Access article distributed in accordance with the Creative Commons Attribution (CC BY 4.0) licence, which permits others to distribute, remix, adapt, build upon this work, and license their derivative works on different terms, provided the original work is properly cited. |
Keywords: | Mhp1; five helix inverted repeat superfamily; hydantoin; membrane transport; molecular recognition; nucleobase‐cation‐symport, NCS1, family; bacterial proteins; binding sites; biological transport; crystallography, x-ray; hydantoins; hydrogen bonding; ligands; micrococcaceae; models, molecular; molecular dynamics simulation; mutagenesis, site-Directed; mutation; protein conformation; structure-activity relationship |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Mechanical Engineering (Leeds) > Institute of Functional Surfaces (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) > Leeds Institute of Cardiovascular and Metabolic Medicine (LICAMM) > Discovery & Translational Science Dept (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 26 Mar 2015 12:18 |
Last Modified: | 20 Jun 2021 08:37 |
Published Version: | http://dx.doi.org/10.15252/embj.201387557 |
Status: | Published |
Publisher: | EMBO Press |
Identification Number: | 10.15252/embj.201387557 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:84088 |