Time-resolved IR spectroscopy reveals mechanistic details of ion transport in the sodium pump Krokinobacter eikastus rhodopsin 2

Phys Chem Chem Phys. 2019 Feb 20;21(8):4461-4471. doi: 10.1039/c8cp07418f.

Abstract

We report a comparative study on the structural dynamics of the light-driven sodium pump Krokinobacter eikastus rhodopsin 2 wild type under sodium and proton pumping conditions by means of time-resolved IR spectroscopy. The kinetics of KR2 under sodium pumping conditions exhibits a sequential character, whereas the kinetics of KR2 under proton pumping conditions involves several equilibrium states. The sodium translocation itself is characterized by major conformational changes of the protein backbone, such as distortions of the α-helices and probably of the ECL1 domain, indicated by distinct marker bands in the amide I region. Carbonyl stretch modes of specific amino acid residues helped to elucidate structural changes in the retinal Schiff base moiety, including the protonation and deprotonation of D116, which is crucial for a deeper understanding of the mechanistic features in the photocycle of KR2.

MeSH terms

  • Cell Membrane / metabolism
  • Escherichia coli / genetics
  • Flavobacteriaceae / metabolism*
  • Flavobacteriaceae / radiation effects
  • Ion Transport
  • Kinetics
  • Light
  • Models, Molecular
  • Molecular Structure
  • Photochemical Processes
  • Rhodopsins, Microbial / metabolism*
  • Rhodopsins, Microbial / radiation effects
  • Sodium Channels / metabolism*
  • Sodium Channels / radiation effects
  • Sodium-Potassium-Exchanging ATPase / metabolism
  • Spectrophotometry, Infrared
  • Thermodynamics

Substances

  • Rhodopsins, Microbial
  • Sodium Channels
  • Sodium-Potassium-Exchanging ATPase