Voltage-clamp fluorometry analysis of structural rearrangements of ATP-gated channel P2X2 upon hyperpolarization

eLife

Rizki Tsari Andriani¹  Yoshihiro Kubo¹ 

[1] Division of Biophysics and Neurobiology, National Institute for Physiological Sciences, Aichi, Japan;Department of Physiological Sciences, The Graduate University for Advanced Studies, School of Life Science, Kanagawa, Japan;

关键词: P2X2;    voltage-clamp fluorometry;    fluorescent unnatural amino acid;    Xenopus;   

DOI  :  10.7554/eLife.65822

来源: eLife Sciences Publications, Ltd

【 摘 要 】

Gating of the ATP-activated channel P2X2 has been shown to be dependent not only on [ATP] but also on membrane voltage, despite the absence of a canonical voltage-sensor domain. We aimed to investigate the structural rearrangements of rat P2X2 during ATP- and voltage-dependent gating, using a voltage-clamp fluorometry technique. We observed fast and linearly voltage-dependent fluorescence intensity (F) changes at Ala337 and Ile341 in the TM2 domain, which could be due to the electrochromic effect, reflecting the presence of a converged electric field. We also observed slow and voltage-dependent F changes at Ala337, which reflect structural rearrangements. Furthermore, we determined that the interaction between Ala337 in TM2 and Phe44 in TM1, which are in close proximity in the ATP-bound open state, is critical for activation. Taking these results together, we propose that the voltage dependence of the interaction within the converged electric field underlies the voltage-dependent gating.

【 授权许可】

CC BY