Back to Search Start Over

Green-Sensitive, Long-Lived, Step-Functional Anion Channelrhodopsin-2 Variant as a High-Potential Neural Silencing Tool.

Green-Sensitive, Long-Lived, Step-Functional Anion Channelrhodopsin-2 Variant as a High-Potential Neural Silencing Tool.

Authors :
Kojima K
Miyoshi N
Shibukawa A
Chowdhury S
Tsujimura M
Noji T
Ishikita H
Yamanaka A
Sudo Y
Source :
The journal of physical chemistry letters [J Phys Chem Lett] 2020 Aug 06; Vol. 11 (15), pp. 6214-6218. Date of Electronic Publication: 2020 Jul 22.
Publication Year :
2020

Abstract

Anion channelrhodopsin-2 (GtACR2) was identified from the alga Guillardia theta as a light-gated anion channel, providing a powerful neural silencing tool for optogenetics. To expand its molecular properties, we produced here GtACR2 variants by strategic mutations on the four residues around the retinal chromophore (i.e., R129, G152, P204, and C233). After the screening with the Escherichia coli expression system, we estimated spectral sensitivities and the anion channeling function by using the HEK293 expression system. Among the mutants, triple (R129M/G152S/C233A) and quadruple (R129M/G152S/P204T/C233A) mutants showed the significantly red-shifted absorption maxima (λ <subscript>max</subscript> = 498 and 514 nm, respectively) and the long-lived channel-conducting states (the half-life times were 3.4 and 5.4 s, respectively). In addition, both mutants can be activated and inactivated by different wavelengths, representing their step-functional ability. We nicknamed the quadruple mutant "GLaS-ACR2" from its green-sensitive, long-lived, step-functional properties. The unique characteristics of GLaS-ACR2 suggest its high potential as a neural silencing tool.

Details

Language :
English
ISSN :
1948-7185
Volume :
11
Issue :
15
Database :
MEDLINE
Journal :
The journal of physical chemistry letters
Publication Type :
Academic Journal
Accession number :
32697087
Full Text :
https://doi.org/10.1021/acs.jpclett.0c01406