Single-molecule dynamics of gating in a neurotransmitter transporter homologue. Academic Article uri icon

Overview

MeSH

  • Alanine
  • Allosteric Regulation
  • Crystallography, X-Ray
  • Cysteine
  • Escherichia coli
  • Fluorescence Resonance Energy Transfer
  • Leucine
  • Models, Molecular
  • Molecular Dynamics Simulation
  • Protein Conformation
  • Sodium

MeSH Major

  • Aquifoliaceae
  • Bacterial Proteins
  • Plasma Membrane Neurotransmitter Transport Proteins

abstract

  • Neurotransmitter:Na(+) symporters (NSS) remove neurotransmitters from the synapse in a reuptake process that is driven by the Na(+) gradient. Drugs that interfere with this reuptake mechanism, such as cocaine and antidepressants, profoundly influence behaviour and mood. To probe the nature of the conformational changes that are associated with substrate binding and transport, we have developed a single-molecule fluorescence imaging assay and combined it with functional and computational studies of the prokaryotic NSS homologue LeuT. Here we show molecular details of the modulation of intracellular gating of LeuT by substrates and inhibitors, as well as by mutations that alter binding, transport or both. Our direct observations of single-molecule transitions, reflecting structural dynamics of the intracellular region of the transporter that might be masked by ensemble averaging or suppressed under crystallographic conditions, are interpreted in the context of an allosteric mechanism that couples ion and substrate binding to transport.

publication date

  • May 13, 2010

has subject area

  • Alanine
  • Allosteric Regulation
  • Aquifoliaceae
  • Bacterial Proteins
  • Crystallography, X-Ray
  • Cysteine
  • Escherichia coli
  • Fluorescence Resonance Energy Transfer
  • Leucine
  • Models, Molecular
  • Molecular Dynamics Simulation
  • Plasma Membrane Neurotransmitter Transport Proteins
  • Protein Conformation
  • Sodium

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC2940119

Digital Object Identifier (DOI)

  • 10.1038/nature09057

PubMed ID

  • 20463731

Additional Document Info

start page

  • 188

end page

  • 193

volume

  • 465

number

  • 7295