Single-molecule transport kinetics of a glutamate transporter homolog shows static disorder

Sci Adv. 2020 May 29;6(22):eaaz1949. doi: 10.1126/sciadv.aaz1949. eCollection 2020 May.

Abstract

Kinetic properties of membrane transporters are typically poorly defined because high-resolution functional assays analogous to single-channel recordings are lacking. Here, we measure single-molecule transport kinetics of a glutamate transporter homolog from Pyrococcus horikoshii, GltPh, using fluorescently labeled periplasmic amino acid binding protein as a fluorescence resonance energy transfer-based sensor. We show that individual transporters can function at rates varying by at least two orders of magnitude that persist for multiple turnovers. A gain-of-function mutant shows increased population of the fast-acting transporters, leading to a 10-fold increase in the mean transport rate. These findings, which are broadly consistent with earlier single-molecule measurements of GltPh conformational dynamics, suggest that GltPh transport is defined by kinetically distinct populations that exhibit long-lasting "molecular memory."

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't