Transport dynamics in a glutamate transporter homologue. Academic Article uri icon

Overview

abstract

  • Glutamate transporters are integral membrane proteins that catalyse neurotransmitter uptake from the synaptic cleft into the cytoplasm of glial cells and neurons. Their mechanism of action involves transitions between extracellular (outward)-facing and intracellular (inward)-facing conformations, whereby substrate binding sites become accessible to either side of the membrane. This process has been proposed to entail transmembrane movements of three discrete transport domains within a trimeric scaffold. Using single-molecule fluorescence resonance energy transfer (smFRET) imaging, we have directly observed large-scale transport domain movements in a bacterial homologue of glutamate transporters. We find that individual transport domains alternate between periods of quiescence and periods of rapid transitions, reminiscent of bursting patterns first recorded in single ion channels using patch-clamp methods. We propose that the switch to the dynamic mode in glutamate transporters is due to separation of the transport domain from the trimeric scaffold, which precedes domain movements across the bilayer. This spontaneous dislodging of the substrate-loaded transport domain is approximately 100-fold slower than subsequent transmembrane movements and may be rate determining in the transport cycle.

publication date

  • June 23, 2013

Research

keywords

  • Amino Acid Transport System X-AG
  • Models, Molecular
  • Pyrococcus horikoshii

Identity

PubMed Central ID

  • PMC3829612

Scopus Document Identifier

  • 84885669136

Digital Object Identifier (DOI)

  • 10.1038/nature12265

PubMed ID

  • 23792560

Additional Document Info

volume

  • 502

issue

  • 7469