The LeuT-fold neurotransmitter:Sodium symporter MhsT has two substrate sites Academic Article uri icon

Overview

MeSH Major

  • Bacillus
  • Bacterial Proteins
  • Molecular Dynamics Simulation
  • Plasma Membrane Neurotransmitter Transport Proteins
  • Sodium

abstract

  • © 2018 National Academy of Sciences. All Rights Reserved. Crystal structures of the neurotransmitter:sodium symporter MhsT revealed occluded inward-facing states with one substrate (Trp) bound in the primary substrate (S1) site and a collapsed extracellular vestibule, which in LeuT contains the second substrate (S2) site. In n-dodecyl-β-D-maltoside, the detergent used to prepare MhsT for crystallization, the substrate-to-protein binding stoichiometry was determined by using scintillation proximity to be 1 Trp:MhsT. Here, using the same experimental approach, as well as equilibrium dialysis, we report that in n-decyl-β-D-maltoside, or after reconstitution in lipid, MhsT, like LeuT, can simultaneously bind two Trp substrate molecules. Trp binding to the S2 site sterically blocks access to a substituted Cys at position 33 in the S2 site, as well as access to the deeper S1 site. Mutation of either the S1 or S2 site disrupts transport, consistent with previous studies in LeuT showing that substrate binding to the S2 site is an essential component of the transport mechanism.

publication date

  • August 21, 2018

Research

keywords

  • Academic Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1073/pnas.1717444115

PubMed ID

  • 30082383

Additional Document Info

start page

  • E7924

end page

  • E7931

volume

  • 115

number

  • 34