Genetically encoded fluorescent voltage sensors using the voltage-sensing domain of Nematostella and Danio phosphatases exhibit fast kinetics

Title
Genetically encoded fluorescent voltage sensors using the voltage-sensing domain of Nematostella and Danio phosphatases exhibit fast kinetics
Authors
브래들리 베이커Lei JinZhou Han래리코헨Marko PopovicJelena PlatisaVincent Pieribone
Keywords
Genetically encoded voltage-sensor; Fluorescent protein; Imaging membrane potential; Voltage-sensing phosphatase
Issue Date
2012-07
Publisher
Journal of neuroscience methods
Citation
VOL 208, 190-196
Abstract
A substantial increase in the speed of the optical response of genetically encoded fluorescent protein voltage sensors (FP voltage sensors) was achieved by using the voltage-sensing phosphatase genes of Nematostella vectensis and Danio rerio. A potential N. vectensis voltage-sensing phosphatase was identified in silico. The voltage-sensing domain (S1&#8211;S4) of the N. vectensis homolog was used to create an FP voltage sensor called Nema. By replacing the phosphatase with a cerulean/citrine FRET pair, a new FP voltage sensor was synthesized with fast off kinetics (Tauoff < 5 ms). However, the signal was small (ΔF/F = 0.4%/200 mV). FP voltage sensors using the D. rerio voltage-sensing phosphatase homolog, designated Zahra and Zahra 2, exhibited fast on and off kinetics within 2 ms of the time constants observed with the organic voltage-sensitive dye, di4-ANEPPS. Mutagenesis of the S4 region of the Danio FP voltage sensor shifted the voltage dependence to more negative potentials but did not noticeably affect the kinetics of the optical signal.
URI
http://pubs.kist.re.kr/handle/201004/46975
ISSN
01650270
Appears in Collections:
KIST Publication > Article
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