Specific residues in the cytoplasmic domain modulate photocurrent kinetics of channelrhodopsin from Klebsormidium nitens

Commun Biol. 2021 Feb 23;4(1):235. doi: 10.1038/s42003-021-01755-5.

Abstract

Channelrhodopsins (ChRs) are light-gated ion channels extensively applied as optogenetics tools for manipulating neuronal activity. All currently known ChRs comprise a large cytoplasmic domain, whose function is elusive. Here, we report the cation channel properties of KnChR, one of the photoreceptors from a filamentous terrestrial alga Klebsormidium nitens, and demonstrate that the cytoplasmic domain of KnChR modulates the ion channel properties. KnChR is constituted of a 7-transmembrane domain forming a channel pore, followed by a C-terminus moiety encoding a peptidoglycan binding domain (FimV). Notably, the channel closure rate was affected by the C-terminus moiety. Truncation of the moiety to various lengths prolonged the channel open lifetime by more than 10-fold. Two Arginine residues (R287 and R291) are crucial for altering the photocurrent kinetics. We propose that electrostatic interaction between the rhodopsin domain and the C-terminus domain accelerates the channel kinetics. Additionally, maximal sensitivity was exhibited at 430 and 460 nm, the former making KnChR one of the most blue-shifted ChRs characterized thus far, serving as a novel prototype for studying the molecular mechanism of color tuning of the ChRs. Furthermore, KnChR would expand the optogenetics tool kit, especially for dual light applications when short-wavelength excitation is required.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Line
  • Channelrhodopsins / chemistry
  • Channelrhodopsins / genetics
  • Channelrhodopsins / metabolism*
  • Channelrhodopsins / radiation effects
  • Chlorophyta / genetics
  • Chlorophyta / metabolism*
  • Chlorophyta / radiation effects
  • Ion Channel Gating* / radiation effects
  • Kinetics
  • Light
  • Membrane Potentials
  • Mice
  • Optogenetics
  • Protein Domains
  • Rats
  • Structure-Activity Relationship

Substances

  • Channelrhodopsins