Bacteriorhodopsin-like channelrhodopsins: Alternative mechanism for control of cation conductance

The recently discovered cation-conducting channelrhodopsins in cryptophyte algae are far more homologous to haloarchaeal rhodopsins, in particular the proton pump bacteriorhodopsin (BR), than to earlier known channelrhodopsins. They uniquely retain the two carboxylate residues that define the vector...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2017-11, Vol.114 (45), p.E9512-E9519
Main Authors: Sineshchekov, Oleg A., Govorunova, Elena G., Li, Hai, Spudich, John L.
Format: Article
Language:English
Subjects:
Algae
Bacteriorhodopsin
Biological Sciences
Cations
Channel gating
Channel opening
Conductance
Cytoplasm
Homology
Imines
Membranes
Mutants
Photochemicals
Pigments
PNAS Plus
Protons
Residues
Resistance
Translocation
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Summary:The recently discovered cation-conducting channelrhodopsins in cryptophyte algae are far more homologous to haloarchaeal rhodopsins, in particular the proton pump bacteriorhodopsin (BR), than to earlier known channelrhodopsins. They uniquely retain the two carboxylate residues that define the vectorial proton path in BR in which Asp-85 and Asp-96 serve as acceptor and donor, respectively, of the photoactive site Schiff base (SB) proton. Here we analyze laser flash-induced photocurrents and photochemical conversions in Guillardia theta cation channelrhodopsin 2 (GtCCR2) and its mutants. Our results reveal a model in which the GtCCR2 retinylidene SB chromophore rapidly deprotonates to the Asp-85 homolog, as in BR. Opening of the cytoplasmic channel to cations in GtCCR2 requires the Asp-96 homolog to be unprotonated, as has been proposed for the BR cytoplasmic channel for protons. However, reprotonation of the GtCCR2 SB occurs not from the Asp-96 homolog, but by proton return from the earlier protonated acceptor, preventing vectorial proton translocation across the membrane. In GtCCR2, deprotonation of the Asp-96 homolog is required for cation channel opening and occurs >10-fold faster than reprotonation of the SB, which temporally correlates with channel closing. Hence in GtCCR2, cation channel gating is tightly coupled to intramolecular proton transfers involving the same residues that define the vectorial proton path in BR.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1710702114