Genetically encoded fluorescent voltage sensors using the voltage-sensing domain of Nematostella and Danio phosphatases exhibit fast kinetics.
Genetically encoded fluorescent voltage sensors using the voltage-sensing domain of Nematostella and Danio phosphatases exhibit fast kinetics.
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DOI：
10.1016/j.jneumeth.2012.05.016
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发表时间：
2012-07-15
影响因子：
3
通讯作者：
Pieribone V
中科院分区：
文献类型：
--
作者：
Baker BJ;Jin L;Han Z;Cohen LB;Popovic M;Platisa J;Pieribone V
利用线虫和丹尼奥韦里奥的电压感应磷酸酶基因，实现了基因编码荧光蛋白电压传感器（FP电压传感器）光响应速度的大幅提高。在硅片上鉴定出一种潜在的维氏乳杆菌电压感应磷酸酶。利用N. vectensis同系物的电压传感域（S1-S4）创建FP电压传感器Nema。用天蓝色/黄嘌呤FRET对取代磷酸酶，合成了具有快速关闭动力学（Tauoff <5 msec）的新型FP电压传感器。但信号很小（ΔF/F= 0.6%/200 mV）。使用D. rerio电压感应磷酸酶同源物的FP电压传感器，命名为Zahra和Zahra 2，在使用有机电压敏感染料di4-ANEPPS观察到的时间常数2毫秒内表现出快速的开关动力学。Danio FP电压传感器S4区域的突变将电压依赖性转移到更多的负电位，但对光信号的动力学没有明显影响。
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–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 msec). However, the signal was small (ΔF/F= 0.6%/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 msec 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.
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影响因子：
3.7
作者：
Dimitrov D;He Y;Mutoh H;Baker BJ;Cohen L;Akemann W;Knöpfel T
通讯作者：
Knöpfel T
影响因子：
3
作者：
Baker, B. J.;Lee, H.;Kosmidis, E. K.
通讯作者：
Kosmidis, E. K.
影响因子：
48
作者：
Tsutsui, Hidekazu;Karasawa, Satoshi;Miyawaki, Atsushi
通讯作者：
Miyawaki, Atsushi
DOI：
10.1038/newbio241159a0
发表时间：
1973-01-01
期刊：
NATURE-NEW BIOLOGY
影响因子：
--
作者：
DAVILA, HV;SALZBERG, BM;WAGGONER, AS
通讯作者：
WAGGONER, AS
影响因子：
5.8
作者：
Larkin, M. A.;Blackshields, G.;Higgins, D. G.
通讯作者：
Higgins, D. G.