Direct Gating of the TRPM2 Channel by cADPR via Specific Interactions with the ADPR Binding Pocket
Direct Gating of the TRPM2 Channel by cADPR via Specific Interactions with the ADPR Binding Pocket
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cADPR 通过与 ADPR 绑定袋的特定交互直接选通 TRPM2 通道
DOI：
10.1016/j.celrep.2019.05.067
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发表时间：
2019
期刊：
影响因子：
8.8
通讯作者：
Yang Wei
中科院分区：
文献类型：
--
作者：
Yu Peilin;Liu Zhenming;Yu Xiafei;Ye Peiwu;Liu Huan;Xue Xiwen;Yang Lixin;Li Zhongtang;Wu Yang;Fang Cheng;Zhao Yong Juan;Yang Fan;Luo Jian Hong;Jiang Lin Hua;Zhang Liangren;Zhang Lihe;Yang Wei
cADPR是一种通过调节RyRs而得到广泛认可的信号分子，但关于cADPR是否可以结合并调控TRPM2通道存在相当大的争议，TRPM2通道在多种生理和病理过程中介导氧化应激信号。在这里，我们发现纯化的cADPR在全细胞和无细胞单通道记录中都诱发了TRPM2通道电流，并且cADPR通过表面等离子体共振与纯化的TRPM2的NUDT9-H结构域特异性结合。此外，通过将计算模型与电生理记录相结合，我们发现携带H1346、T1347、L1379、S1391、E1409和L1484点突变的TRPM2通道对ADPR和cADPR具有不同的敏感性。这些结果清楚地表明cADPR是TRPM2通道上的非同源失活因子，并清楚地描绘了cADPR结合的结构基础，这不仅有助于更好地理解TRPM2通道的门控机制，而且有助于揭示cADPR诱导的不依赖于ryrs的Ca2+信号传导机制。
cADPR is a well-recognized signaling molecule by modulating the RyRs, but considerable debate exists regarding whether cADPR can bind to and gate the TRPM2 channel, which mediates oxidative stress signaling in diverse physiological and pathological processes. Here, we show that purified cADPR evoked TRPM2 channel currents in both whole-cell and cell-free single-channel recordings and specific binding of cADPR to the purified NUDT9-H domain of TRPM2 by surface plasmon resonance. Furthermore, by combining computational modeling with electrophysiological recordings, we show that the TRPM2 channels carrying point mutations at H1346, T1347, L1379, S1391, E1409, and L1484 possess distinct sensitivity profiles for ADPR and cADPR. These results clearly indicate cADPR is abona fideactivator at the TRPM2 channel and clearly delineate the structural basis for cADPR binding, which not only lead to a better understanding in the gating mechanism of TRPM2 channel but also shed light on a cADPR-induced RyRs-independent Ca2+signaling mechanism.