Photoactivation of Channelrhodopsin*

Channelrhodopsins (ChRs) are light-gated ion channels that control photomovement of microalgae. In optogenetics, ChRs are widely applied for light-triggering action potentials in cells, tissues, and living animals, yet the spectral properties and photocycle of ChR remain obscure. In this study, we cloned a ChR from the colonial alga Volvox carteri, VChR. After electrophysiological characterization in Xenopus oocytes, VChR was expressed in COS-1 cells and purified. Time-resolved UV-visible spectroscopy revealed a pH-dependent equilibrium of two dark species, D470/D480. Laser flashes converted both with τ ≈ 200 μs into major photointermediates P510/P530, which reverted back to the dark states with τ ≈ 15-100 ms. Both intermediates were assigned to conducting states. Three early intermediates P500/P515 and P390 were detected on a ns to μs time scale. The spectroscopic and electrical data were unified in a photocycle model. The functional expression of VChR we report here paves the way toward a broader structure/function analysis of the recently identified class of light-gated ion channels.

• Publication year

  2008

• Venue

  Journal of Biological Chemistry

• Publication date

  2008-01-18

• Fields of study

  Biology, Medicine, Chemistry, Environmental Science

• Identifiers
  DOI 10.1074/jbc.M708039200PMID 17993465
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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