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

Abstract:

: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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Lee EEL,Bezanilla F

doi

10.1016/j.bpj.2017.10.018

subject

Has Abstract

pub_date

2017-11-21 00:00:00

pages

2178-2181

issue

10

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(17)31133-5

journal_volume

113

pub_type

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