Molecular mechanism of ligand recognition by a membrane transport protein, Mhp1. Simmons, K. J., Jackson, S. M., Brueckner, F., Patching, S. G., Beckstein, O., Ivanova, E., Geng, T., Weyand, S., Drew, D., Lanigan, J., Sharples, D. J., Sansom, M. S., Iwata, S., Fishwick, C. W., Johnson, A. P., Cameron, A. D., & Henderson, P. J. The EMBO Journal, 33:1831–1844, June, 2014.
Molecular mechanism of ligand recognition by a membrane transport protein, Mhp1 [link]Paper  doi  abstract   bibtex   
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. Synopsis Structure‐function and molecular dynamics analysis of the hydantoin active transporter Mhp1 yields a novel intermediate state and delineates the basis for substrate specificity and membrane transport. Hydantoin substrates like indolylmethylhydantoin (IMH) bind to the LeuT‐like Mhp1 transporter in an extended conformationSelectivity of Mhp1 for the substrate is conferred by hydrogen bonds to the hydantoin moiety and the fit of aromatic substituent into a hydrophobic pocketNaphthylmethylhydantoin (NMH) inhibits Mhp1 but is not transportedCrystal structure of Mhp1 with NMH shows TMH10 to adopt the position seen in the outward‐open rather than the occluded state.Mutation of Leu363Ala in TMH10 of Mhp1 converts NMH from an inhibitor to a substrate.
@article{simmons_molecular_2014,
	title = {Molecular mechanism of ligand recognition by a membrane transport protein, {Mhp1}},
	volume = {33},
	copyright = {© 2014 The Authors. Published under the terms of the CC BY 4.0 license. This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.},
	issn = {0261-4189, 1460-2075},
	url = {http://emboj.embopress.org/content/33/16/1831},
	doi = {10.15252/embj.201387557},
	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.
Synopsis



Structure‐function and molecular dynamics analysis of the hydantoin active transporter Mhp1 yields a novel intermediate state and delineates the basis for substrate specificity and membrane transport.

Hydantoin substrates like indolylmethylhydantoin (IMH) bind to the LeuT‐like Mhp1 transporter in an extended conformationSelectivity of Mhp1 for the substrate is conferred by hydrogen bonds to the hydantoin moiety and the fit of aromatic substituent into a hydrophobic pocketNaphthylmethylhydantoin (NMH) inhibits Mhp1 but is not transportedCrystal structure of Mhp1 with NMH shows TMH10 to adopt the position seen in the outward‐open rather than the occluded state.Mutation of Leu363Ala in TMH10 of Mhp1 converts NMH from an inhibitor to a substrate.},
	language = {en},
	urldate = {2014-06-25},
	journal = {The EMBO Journal},
	author = {Simmons, Katie J. and Jackson, Scott M. and Brueckner, Florian and Patching, Simon G. and Beckstein, Oliver and Ivanova, Ekaterina and Geng, Tian and Weyand, Simone and Drew, David and Lanigan, Joseph and Sharples, David J. and Sansom, Mark SP and Iwata, So and Fishwick, Colin WG and Johnson, A. Peter and Cameron, Alexander D. and Henderson, Peter JF},
	month = jun,
	year = {2014},
	keywords = {Mhp1, five helix inverted repeat superfamily, hydantoin, membrane transport, molecular recognition, nucleobase‐cation‐symport, NCS1, family},
	pages = {1831--1844},
}
Downloads: 0
About
Documentation
Keywords
Search
Users
Terms and Conditions of Use
Privacy Policy
Sitemap
© BibBase.org 2021