CLIC2-RyR1 interaction and structural characterization by cryo-electron microscopy.
CLIC2-RyR1 interaction and structural characterization by cryo-electron microscopy.
复制标题
通过冷冻电子显微镜观察 CLIC2-RyR1 相互作用和结构表征。
DOI：
10.1016/j.jmb.2009.01.059
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发表时间：
2009-03-27
影响因子：
5.6
通讯作者：
Yin, Chang-Cheng
中科院分区：
文献类型：
--
作者：
Meng, Xing;Wang, Guoliang;Viero, Cedric;Wang, Qiongling;Mi, Wei;Su, Xiao-Dong;Wagenknecht, Terence;Williams, Alan J.;Liu, Zheng;Yin, Chang-Cheng
关键词：
氯离子胞内通道2（CLIC 2）是一种新发现的小分子蛋白，与谷胱甘肽转移酶（GST）结构家族有较远的亲缘关系，在心肌和骨骼肌中高度表达，但其在这些组织中的生理功能尚未确定。在本研究中，[3 H]-ryanodine结合，Ca 2+流出骨骼肌肌浆网（SR）囊泡，单通道记录，和冷冻电镜研究CLIC 2是否可以与骨骼肌ryanodine受体（RyR 1）相互作用，并调节其通道活性。我们发现：（1）CLIC 2通过增加ryanodine与骨骼肌SR和纯化RyR 1的结合亲和力而促进[3 H]-ryanodine与其受体的结合，但不显著改变表观最大结合容量，（2）CLIC 2降低骨骼肌SR囊泡的最大Ca ~（2+）流出率，（3）CLIC 2通过增加通道的平均关闭时间而降低RyR 1通道的开放概率，（4）CLIC 2通过增加通道的平均关闭时间而降低通道的开放概率，（5）CLIC 2通过增加通道的平均关闭时间而降低通道的开放概率，（6）CLIC 2通过增加通道的平均关闭时间而降低通道的开放概率，（7）CLIC 2通过增加通道的平均关闭时间而降低通道的开放概率。（4）CLIC 2与RyR 1的钳形区域中的结构域5和结构域6之间的区域结合;（5）在同一钳形区域中，结构域9和结构域10在CLIC 2结合后分离，表明CLIC 2诱导RyR 1的构象变化。这些数据表明CLIC 2可以与RyR 1相互作用并调节其通道活性。我们建议CLIC 2作为RyR通道闭合状态的内在稳定剂发挥作用。
Chloride intracellular channel 2 (CLIC2), a newly discovered small protein distantly related to the glutathione transferase (GST) structural family, is highly expressed in cardiac and skeletal muscle although its physiological function in these tissues has not been established. In the present study, [3H]-ryanodine binding, Ca2+ efflux from skeletal sarcoplasmic reticulum (SR) vesicles, single channel recording, and cryo-electron microscopy were employed to investigate whether CLIC2 can interact with skeletal ryanodine receptor (RyR1) and modulate its channel activity. We found that: (1) CLIC2 facilitated [3H]-ryanodine binding to skeletal SR and purified RyR1, by increasing the binding affinity of ryanodine for its receptor without significantly changing the apparent maximal binding capacity; (2) CLIC2 reduced the maximal Ca2+ efflux rate from skeletal SR vesicles; (3) CLIC2 decreased the open probability of RyR1 channel, through increasing the mean closed time of the channel; (4) CLIC2 bound to a region between domain 5 and domain 6 in the clamp-shaped region of RyR1; (5) and in the same clamp region, domain 9 and domain 10 became separated after CLIC2 binding, indicating CLIC2 induced a conformational change of RyR1. These data suggest that CLIC2 can interact with RyR1 and modulate its channel activity. We propose that CLIC2 functions as an intrinsic stabilizer of the closed state of RyR channels.
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DOI：
10.1152/ajprenal.1999.276.3.f398
发表时间：
1999-03-01
影响因子：
4.2
作者：
Edwards, JC
通讯作者：
Edwards, JC
影响因子：
4.8
作者：
Harrop, SJ;DeMaere, MZ;Curmi, PMG
通讯作者：
Curmi, PMG
影响因子：
64.5
作者：
BRILLANTES, AMB;ONDRIAS, K;MARKS, AR
通讯作者：
MARKS, AR
影响因子：
4.8
作者：
Gaburjakova, M;Gaburjakova, J;Marks, AR
通讯作者：
Marks, AR
影响因子：
3.3
作者：
Berryman, M;Bretscher, A
通讯作者：
Bretscher, A