Li Haiwen, Xu Li, Gao Yandi, Zuo Yuanbojiao, Yang Zuocheng, Zhao Lingling, Chen Zhiheng, Guo Shuliang, Han Renzhi
Division of Cardiac Surgery, Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, 43210, USA.
Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, China.
Cell Biosci. 2021 Dec 28;11(1):222. doi: 10.1186/s13578-021-00735-w.
Anoctamin 5 (ANO5) is a membrane protein belonging to the TMEM16/Anoctamin family and its deficiency leads to the development of limb girdle muscular dystrophy R12 (LGMDR12). However, little has been known about the interactome of ANO5 and its cellular functions.
In this study, we exploited a proximal labeling approach to identify the interacting proteins of ANO5 in C2C12 myoblasts stably expressing ANO5 tagged with BioID2. Mass spectrometry identified 41 unique proteins including BVES and POPDC3 specifically from ANO5-BioID2 samples, but not from BioID2 fused with ANO6 or MG53. The interaction between ANO5 and BVES was further confirmed by co-immunoprecipitation (Co-IP), and the N-terminus of ANO5 mediated the interaction with the C-terminus of BVES. ANO5 and BVES were co-localized in muscle cells and enriched at the endoplasmic reticulum (ER) membrane. Genome editing-mediated ANO5 or BVES disruption significantly suppressed C2C12 myoblast differentiation with little impact on proliferation.
Taken together, these data suggest that BVES is a novel interacting protein of ANO5, involved in regulation of muscle differentiation.
anoctamin 5（ANO5）是一种属于TMEM16/anoctamin家族的膜蛋白，其缺陷会导致肢带型肌营养不良R12（LGMDR12）的发生。然而，关于ANO5的相互作用组及其细胞功能知之甚少。
在本研究中，我们采用近端标记方法，在稳定表达用BioID2标记的ANO5的C2C12成肌细胞中鉴定ANO5的相互作用蛋白。质谱分析从ANO5-BioID2样品中鉴定出41种独特的蛋白质，包括BVES和POPDC3，但在与ANO6或MG53融合的BioID2样品中未鉴定到。通过免疫共沉淀（Co-IP）进一步证实了ANO5与BVES之间的相互作用，ANO5的N端介导了与BVES C端的相互作用。ANO5和BVES在肌肉细胞中共定位，并在内质网（ER）膜上富集。基因组编辑介导的ANO5或BVES破坏显著抑制C2C12成肌细胞分化，对增殖影响很小。
综上所述，这些数据表明BVES是ANO5的一种新型相互作用蛋白，参与肌肉分化的调节。
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