Yeast two-hybrid screening identifies binding partners of human Tom34 that have ATPase activity and form a complex with Tom34 in the cytosol.
Yeast two-hybrid screening identifies binding partners of human Tom34 that have ATPase activity and form a complex with Tom34 in the cytosol.
复制标题
酵母双杂交筛选鉴定出具有 ATP 酶活性并与细胞质中的 Tom34 形成复合物的人 Tom34 的结合伴侣。
DOI：
10.1006/abbi.2002.2778
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Weiner,Henry
中科院分区：
文献类型：
--
作者：
Yang,Chun-Song;Weiner,Henry
附文(MukHopadhyay，A.，Avramova，L.V.和Weiner，H.，Arch.生物化学。生物物理研究表明，Tom34是一种先前提出的线粒体外膜转位酶，它与前体蛋白的成熟区结合，似乎是一种胞浆蛋白。以Tom34为诱饵进行酵母双杂交筛选，寻找其潜在的结合伙伴。其中两个被鉴定的蛋白质是与ATPase相关的Valosin蛋白(VCP)和溶酶体H+转运ATPase成员M(ATP6M)。Tom34主要分布在细胞质中，VCP和ATP6M主要分布在细胞质和非线粒体细胞器中。Tom34与它们在胞浆中形成∼400 kDa的复合体。发现Tom34具有弱的ATPase活性，当与VCP结合时，该活性不会改变。Tom34的四肽重复(TPR)基序区域(残基201-256)负责与其他蛋白质结合。Tom34似乎不是线粒体外膜转位酶家族的成员，但在蛋白转位过程中可能作为一种伴侣样蛋白发挥作用。
In the accompany paper (Mukhopadhyay, A., Avramova, L. V. and Weiner, H., Arch. Biochem. Biophys.), it was shown that Tom34, a previously proposed putative translocase of the mitochondrial outer membrane, binds to the mature region of a precursor protein and appears to be a cytosol protein. Here Tom34 was used as bait in a yeast two-hybrid screening to search for its potential binding partners. Two of the identified proteins were the ATPase-related valosin-containing protein (VCP) and the lysosomal H+-transporting ATPase member M (ATP6M). Tom34 was found primarily in the cytosol while VCP and ATP6M were found in the cytosol as well as in nonmitochondrial organelles. Tom34 formed a ∼400-kDa complex with them in the cytosol. Tom34 was found to possess a weak ATPase activity that did not change when associated with VCP. The tetratricopeptide repeat (TPR) motif region of Tom34 (residue 201–256) was responsible for binding to the other proteins. Tom34 appears not to be a member of the mitochondrial outer membrane translocase family but might function as a chaperone-like protein during protein translocation.