Abstract：
BACKGROUND: 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.
RESULTS: 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.
CONCLUSIONS: Taken together, these data suggest that BVES is a novel interacting protein of ANO5, involved in regulation of muscle differentiation.
RESULTS: 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.
CONCLUSIONS: Taken together, these data suggest that BVES is a novel interacting protein of ANO5, involved in regulation of muscle differentiation.
摘要：
背景：Anoctamin5（ANO5）是属于TMEM16/Anoctamin家族的膜蛋白，其缺乏导致肢带肌营养不良R12（LGMDR12）的发展。然而,关于ANO5的相互作用组及其细胞功能知之甚少。
结果：在这项研究中，我们利用近端标记方法来鉴定稳定表达用BioID2标记的ANO5的C2C12成肌细胞中ANO5的相互作用蛋白。质谱从ANO5-BioID2样品中鉴定出41种独特的蛋白质，包括BVES和POPDC3，但不是来自与ANO6或MG53融合的BioID2。通过免疫共沉淀(Co-IP)进一步证实了ANO5和BVES之间的相互作用,ANO5的N端介导与BVES的C端相互作用。ANO5和BVES共定位在肌细胞中并富集在内质网(ER)膜。基因组编辑介导的ANO5或BVES破坏显着抑制C2C12成肌细胞分化，对增殖影响很小。
结论：综合来看，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。
结果：在这项研究中，我们利用近端标记方法来鉴定稳定表达用BioID2标记的ANO5的C2C12成肌细胞中ANO5的相互作用蛋白。质谱从ANO5-BioID2样品中鉴定出41种独特的蛋白质，包括BVES和POPDC3，但不是来自与ANO6或MG53融合的BioID2。通过免疫共沉淀(Co-IP)进一步证实了ANO5和BVES之间的相互作用,ANO5的N端介导与BVES的C端相互作用。ANO5和BVES共定位在肌细胞中并富集在内质网(ER)膜。基因组编辑介导的ANO5或BVES破坏显着抑制C2C12成肌细胞分化，对增殖影响很小。
结论：综合来看，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。
