Global Symmetry: Asymmetric - C1
Global Stoichiometry: Monomer - A
Biological assembly 1 assigned by authors and generated by PISA (software)
4D1A
STRUCTURE OF MHP1, A NUCLEOBASE-CATION-SYMPORT-1 FAMILY TRANSPORTER, IN A CLOSED CONFORMATION WITH INDOLYLMETHYL-HYDANTOIN
DOI: 10.2210/pdb4D1A/pdb
Classification: TRANSPORT PROTEIN
Organism(s): Microbacterium liquefaciens
Expression System: Escherichia coli BL21(DE3)
Deposition Author(s): Weyand, S., Brueckner, F., Geng, T., Drew, D., Iwata, S., Henderson, P.J.F., Cameron, A.D.
Molecular Mechanism of Ligand Recognition by 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.
(2014) Embo J. 33: 1831
PubMed: 24952894 Search on PubMedSearch on PubMed Central
DOI: 10.15252/embj.201387557
4D1D, 4D1C, 4D1B
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 sol ...
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.
School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.,School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK p.j.f.henderson@leeds.ac.uk a.cameron@warwick.ac.uk c.w.g.fishwick@leeds.ac.uk.,Membrane Protein Laboratory, Diamond Light Source Harwell Science and Innovation Campus, Chilton, Didcot, UK Division of Molecular Biosciences, Membrane Protein Crystallography Group Imperial College, London, UK School of Life Sciences, University of Warwick, Coventry, UK p.j.f.henderson@leeds.ac.uk a.cameron@warwick.ac.uk c.w.g.fishwick@leeds.ac.uk.,School of Biomedical Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK p.j.f.henderson@leeds.ac.uk a.cameron@warwick.ac.uk c.w.g.fishwick@leeds.ac.uk.,Division of Molecular Biosciences, Membrane Protein Crystallography Group Imperial College, London, UK.,School of Biomedical Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.,Membrane Protein Laboratory, Diamond Light Source Harwell Science and Innovation Campus, Chilton, Didcot, UK Division of Molecular Biosciences, Membrane Protein Crystallography Group Imperial College, London, UK Rutherford Appleton Laboratory, Research Complex at Harwell, Harwell, Oxford, Didcot, UK.,Department of Biochemistry, University of Oxford, Oxford, UK.,Department of Physics, Arizona State University, Tempe, AZ, USA Department of Biochemistry, University of Oxford, Oxford, UK.
HYDANTOIN TRANSPORT PROTEIN
495 Microbacterium liquefaciens Mutation(s): 0
Gene Names: hyuP
Find proteins for D6R8X8 (Microbacterium liquefaciens)
Go to UniProtKB: D6R8X8
Full Protein Feature View for D6R8X8
Query on NA
SODIUM ION
FKNQFGJONOIPTF-UHFFFAOYSA-N
I5H
Query on I5H
(5S)-5-(1H-indol-3-ylmethyl)imidazolidine-2,4-dione
C12 H11 N3 O2
RUUREKIGAKIKIL-JTQLQIEISA-N
Space Group: P 21 21 21
a = 87.430 α = 90.00
c = 100.550 γ = 90.00
PHENIX refinement
XSCALE data scaling
PHASER phasing
Type: Database references