Millisecond dynamics of an unlabeled amino acid transporter. Academic Article uri icon

Overview

abstract

  • Excitatory amino acid transporters (EAATs) are important in many physiological processes and crucial for the removal of excitatory amino acids from the synaptic cleft. Here, we develop and apply high-speed atomic force microscopy line-scanning (HS-AFM-LS) combined with automated state assignment and transition analysis for the determination of transport dynamics of unlabeled membrane-reconstituted GltPh, a prokaryotic EAAT homologue, with millisecond temporal resolution. We find that GltPh transporters can operate much faster than previously reported, with state dwell-times in the 50 ms range, and report the kinetics of an intermediate transport state with height between the outward- and inward-facing states. Transport domains stochastically probe transmembrane motion, and reversible unsuccessful excursions to the intermediate state occur. The presented approach and analysis methodology are generally applicable to study transporter kinetics at system-relevant temporal resolution.

publication date

  • October 6, 2020

Research

keywords

  • Amino Acid Transport Systems
  • Image Processing, Computer-Assisted
  • Microscopy, Atomic Force

Identity

PubMed Central ID

  • PMC7538599

Scopus Document Identifier

  • 85092032283

Digital Object Identifier (DOI)

  • 10.1038/s41467-020-18811-z

PubMed ID

  • 33024106

Additional Document Info

volume

  • 11

issue

  • 1