Full Text:   <1901>

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CLC number: Q615

On-line Access: 2019-11-21

Received: 2019-08-09

Revision Accepted: 2019-10-09

Crosschecked: 2019-10-23

Cited: 0

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Citations:  Bibtex RefMan EndNote GB/T7714


Yu-Huan Luo


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Journal of Zhejiang University SCIENCE B 2019 Vol.20 No.12 P.972-982


Effects of calcium-binding sites in the S2–S3 loop on human and Nematostella vectensis TRPM2 channel gating processes

Author(s):  Yu-Huan Luo, Xia-Fei Yu, Cheng Ma, Fan Yang, Wei Yang

Affiliation(s):  Department of Biophysics, Institute of Neuroscience, NHC and CAMS Key Laboratory of Medical Neurobiology, School of Medicine, Zhejiang University, Hangzhou 310058, China; more

Corresponding email(s):   yangwei@zju.edu.cn

Key Words:  TRPM2, Calcium-binding site, S2–, S3 loop, Channel activation, Channel inactivation

Yu-Huan Luo, Xia-Fei Yu, Cheng Ma, Fan Yang, Wei Yang. Effects of calcium-binding sites in the S2–S3 loop on human and Nematostella vectensis TRPM2 channel gating processes[J]. Journal of Zhejiang University Science B, 2019, 20(12): 972-982.

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author="Yu-Huan Luo, Xia-Fei Yu, Cheng Ma, Fan Yang, Wei Yang",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Effects of calcium-binding sites in the S2–S3 loop on human and Nematostella vectensis TRPM2 channel gating processes
%A Yu-Huan Luo
%A Xia-Fei Yu
%A Cheng Ma
%A Fan Yang
%A Wei Yang
%J Journal of Zhejiang University SCIENCE B
%V 20
%N 12
%P 972-982
%@ 1673-1581
%D 2019
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1900477

T1 - Effects of calcium-binding sites in the S2–S3 loop on human and Nematostella vectensis TRPM2 channel gating processes
A1 - Yu-Huan Luo
A1 - Xia-Fei Yu
A1 - Cheng Ma
A1 - Fan Yang
A1 - Wei Yang
J0 - Journal of Zhejiang University Science B
VL - 20
IS - 12
SP - 972
EP - 982
%@ 1673-1581
Y1 - 2019
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1900477

As a crucial signaling molecule, calcium plays a critical role in many physiological and pathological processes by regulating ion channel activity. Recently, one study resolved the structure of the transient receptor potential melastatin 2 (TRPM2) channel from Nematostella vectensis (nvTRPM2). This identified a calcium-binding site in the s2–;s3 loop, while its effect on channel gating remains unclear. Here, we investigated the role of this calcium-binding site in both nvTRPM2 and human TRPM2 (hTRPM2) by mutagenesis and patch-clamp recording. Unlike hTRPM2, nvTRPM2 cannot be activated by calcium alone. Moreover, the inactivation rate of nvTRPM2 was decreased as intracellular calcium concentration was increased. In addition, our results showed that the four key residues in the calcium-binding site of s2–;s3 loop have similar effects on the gating processes of nvTRPM2 and hTRPM2. Among them, the mutations at negatively charged residues (glutamate and aspartate) substantially decreased the currents of nvTRPM2 and hTRPM2. This suggests that these sites are essential for calcium-dependent channel gating. For the charge-neutralizing residues (glutamine and asparagine) in the calcium-binding site, our data showed that glutamine mutating to alanine or glutamate did not affect the channel activity, but glutamine mutating to lysine caused loss of function. Asparagine mutating to aspartate still remained functional, while asparagine mutating to alanine or lysine led to little channel activity. These results suggest that the side chain of glutamine has a less contribution to channel gating than does asparagine. However, our data indicated that both glutamine mutating to alanine or glutamate and asparagine mutating to aspartate accelerated the channel inactivation rate, suggesting that the calcium-binding site in the s2–;s3 loop is important for calcium-dependent channel inactivation. Taken together, our results uncovered the effect of four key residues in the s2–;s3 loop of TRPM2 on the TRPM2 gating process.

S2-S3 loop中钙离子结合位点对人类及海葵来源TRPM2通道门控过程的影响研究

目的:揭示S2-S3 loop的钙离子结合位点对M2型瞬时受体电位通道(TRPM2)门控过程的影响.
创新点:首次探究了人类TRPM2通道(hTRPM2)和海葵TRPM2通道(nvTRPM2)S2-S3 loop的钙离子结合位点内四个氨基酸残基不同突变对通道激活及失活过程的影响,明确了钙离子结合位点对通道门控的作用.

关键词:M2型瞬时受体电位通道(TRPM2);钙离子结合位点;S2-S3 loop;通道激活;通道失活

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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