Biophysical Characterization of Genetically Encoded Voltage Sensor ASAP1: Dynamic Range Improvement

被引:9
作者
Lee, Elizabeth E. L. [1 ]
Bezanilla, Francisco [1 ,2 ,3 ]
机构
[1] Univ Chicago, Comm Neurobiol, Chicago, IL 60637 USA
[2] Univ Chicago, Dept Biochem & Mol Biol, 920 E 58Th St, Chicago, IL 60637 USA
[3] Univ Chicago, Inst Biophys Dynam, Chicago, IL 60637 USA
基金
美国国家卫生研究院;
关键词
FLUORESCENT PROTEIN; SENSING DOMAIN; NEURONS; PROBE;
D O I
10.1016/j.bpj.2017.10.018
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Recent work has introduced a new fluorescent voltage sensor, ASAP1, which can monitor rapid trains of action potentials in cultured neurons. This indicator is based on the Gallus gallus voltage-sensitive phosphatase with the phosphatase domain removed and a circularly permuted GFP placed in the S3-S4 linker. However, many of the biophysical details of this indicator remain unknown. In this work, we study the biophysical properties of ASAP1. Using the cut-open voltage clamp technique, we have simultaneously recorded fluorescence signals and gating currents from Xenopus laevis oocytes expressing ASAP1. Gating charge movement and fluorescence kinetics track closely with each other, although ASAP1 gating currents are significantly faster than those of Ciona intestinalis voltage-sensitive phosphatase. Altering the residue before the first gating charge removes a split in the ASAP1 QV curve, but preserves the accelerated kinetics that allow for the faithful tracking of action potentials in neurons.
引用
收藏
页码:2178 / 2181
页数:4
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