Association of human transferrin receptor with GABARAP
Association of human transferrin receptor with GABARAP
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DOI：
10.1016/s0014-5793(02)02655-8
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发表时间：
2002-05-08
期刊：
影响因子：
3.5
通讯作者：
Enns, CA
中科院分区：
文献类型：
--
作者：
Green, F;O'Hare, T;Enns, CA
酵母双杂交筛选发现转铁蛋白受体(TFR)的细胞质结构域与GABARAP之间存在特异的相互作用，GABARAP是一种14 kDa的蛋白质，与神经元GABA(A)受体的Gamma2亚单位结合。TFR-GABARAP相互作用的特异性通过与纯化蛋白的体外结合试验和GABARAP与HeLa细胞裂解物中内源性TFR的免疫共沉淀来证明。在酵母双杂交筛选和体外结合试验中，在TFR的细胞质区域内用ATRA替换YTRF内化基序减少了与GABARAP的相互作用。利用嵌合GABARAP-GFP对GABARAP进行细胞内定位，结果表明GABARAP主要定位于核周小泡中。我们的结果表明，GABARAP在神经细胞和GABAA受体之外发挥着更广泛的作用。(C)2002年，由Elsevier Science B.V.代表欧洲生化学会联合会出版。
A yeast two-hybrid screen identified a specific interaction between the cytoplasmic domain of transferrin receptor (TfR) and GABARAP, a 14 kDa protein that binds to the gamma2 subunit of neuronal GABA(A) receptors. The specificity of the TfR-GABARAP interaction was demonstrated by in vitro binding assays with purified proteins and by co-immunoprecipitation of GABARAP with endogenous TfR from HeLa cell lysates. Replacement of the YTRF internalization motif with ATRA within the cytoplasmic domain of TfR reduced interaction with GABARAP in the yeast two-hybrid screen and in vitro binding assays. The intracellular location of GABARAP using chimeric GABARAP-GFP showed that the majority of GABARAP was located in perinuclear vesicles. Our results show that GABARAP plays a more general role outside the confines of neuronal cells and GABAA receptors. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.