Neural-specific distribution of transmembrane protein TMEM240 and formation of TMEM240-Body
Neural-specific distribution of transmembrane protein TMEM240 and formation of TMEM240-Body
复制标题
跨膜蛋白 TMEM240 的神经特异性分布和 TMEM240-Body 的形成
DOI：
10.1016/j.ijbiomac.2020.06.080
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发表时间：
2020
影响因子：
8.2
通讯作者：
Jinghua Wang
中科院分区：
文献类型：
--
作者：
Qiongqiong Hu;Guangyou Wang;Xin Chen;Liulei Zhang;Wei Zhao;Yan Jiang;Chong Zhang;Jin Sun;Hao Xu;Hulun Li;Qingfei Kong;Jiarui Zhao;Xinrong Li;Xiaoyu Zhang;Weiqi Lv;Yumei Liu;Gaiqing Yang;Lili Mu;Jinghua Wang
TMEM240的突变被认为是导致SCA21的原因，但具体机制尚未阐明。 TMEM240的亚细胞定位、具体生物学功能和相应的作用机制也尚未被阐明。在本研究中，分别使用 qPCR 和蛋白质印迹评估 TMEM240 的 mRNA 和蛋白表达。利用活细胞成像确定TMEM240的亚细胞位置，并利用电子显微镜确定TMEM240在细胞中的形态和分布。 TMEM240 在神经元中特异性表达。外源TMEM240形成多层细胞结构，我们将其称为TMEM240-Body（T240-Body）。通过离心和过滤分离和纯化T240-Body。 Ni-NTA 琼脂糖上的锚定蛋白 His-tagged-GFP-BP 用于拉下 T240-GFP 结合蛋白。 TMEM240的N端和C端均被证实位于T240-Body内部。共定位实验表明，过氧化物酶体可能有助于 T240-Body 的形成，并且 TMEM240 的两个跨膜区域似乎对于 T240-Body 的形成至关重要。当与 TMEM240 结合时，Emerin 蛋白有助于 T240-Body 的形成。总体而言，这项研究提供了对 TMEM240 的新见解，为未来的研究提供了信息，以进一步了解其生物学功能。
Mutation in TMEM240 is suggested to cause SCA21, but the specific mechanism has not been clarified. The subcellular localization, specific biological function, and corresponding mechanism of action of TMEM240 have also not been delineated. In this study, the mRNA and protein expression of TMEM240 were assessed using qPCR and western blotting, respectively. Live cell imaging was used to establish the sub-cellular location of TMEM240, and electron microscopy was used to determine the morphology and distribution of TMEM240 in the cell. TMEM240 was specifically expressed in the neurons. Exogenous TMEM240 formed a multilayered cell structure, which we refer to as TMEM240-Body (T240-Body). T240-Body was separated and purified by centrifugation and filtration. An anchor protein His-tagged-GFP-BP on Ni-NTA agarose was used to pull down T240-GFP binding proteins. Both the N-terminal and the C-terminal of TMEM240 were confirmed to be inside the T240-Body. Co-localization experiments suggested that peroxisomes might contribute to T240-Body formation, and the two transmembrane regions of TMEM240 appear to be essential for formation of the T240-Body. Emerin protein contributed to formation of T240-Body when combined with TMEM240. Overall, this study provides new insights into TMEM240, which inform future research to further our understanding of its biological function.