CLIC2-RyR1 Interaction and Structural Characterization by Cryo-electron Microscopy

JOURNAL OF MOLECULAR BIOLOGY

卷:387

Article

Meng, Xing²  Wang, Guoliang³  Viero, Cedric¹  Wang, Qiongling³  Mi, Wei⁴  Su, Xiao-Dong⁴  Wagenknecht, Terence²  Williams, Alan J.¹  Liu, Zheng²  Yin, Chang-Cheng³ 

[1] Cardiff Univ, Sch Med, Wales Heart Res Inst, Dept Cardiol, Cardiff CF14 4XN, S Glam, Wales

[2] New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA

[3] Peking Univ, Peking Univ Hlth Sci Ctr, Dept Biophys, Beijing 100191, Peoples R China

[4] Peking Univ, Coll Life Sci, Natl Lab Prot Engn & Plant Genet Engn, Beijing 100871, Peoples R China

关键词: Ca2+-release channel;    Ca2+ signaling;    chloride intracellular channel 2;    cryo-electron microscopy;    ryanodine receptor;   

DOI  :  10.1016/j.jmb.2009.01.059

来源: Elsevier

【 摘 要 】

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, [H-3] 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 [H-3]ryanodine binding to skeletal SR and purified RyR1, by increasing the binding affinity of ryanodine for its receptor without significantly changing the a parent 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 domains 5 and 6 in the clamp-shaped region of RyR1; (5) and in the same clamp region, domains 9 and 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. (C) 2009 Elsevier Ltd. All rights reserved.

【 授权许可】

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