Analysis of proteins interacting with TRIP8b adapter
Analysis of proteins interacting with TRIP8b adapter
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DOI：
10.1134/s0006297908060035
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发表时间：
2008-06-01
影响因子：
2.8
通讯作者：
Petrenko, A. G.
中科院分区：
文献类型：
--
作者：
Popova, N. V.;Plotnikov, A. N.;Petrenko, A. G.
蛛毒素钙非依赖性受体 (CIRL) 是一种孤儿七螺旋受体，参与胞吐作用的调节。为了表征 CIRL 功能的分子机制，我们使用酵母双杂交 SR 系统，以 CIRL 的细胞质 C 端片段作为诱饵，寻找其细胞内伙伴。其中一个相互作用的蛋白被鉴定为 TRIP8b，一种假定的具有多个四三肽重复序列的胞质接头蛋白。为了了解 CIRL-TRIP8b 相互作用的功能意义，我们通过固定化重组 TRIP8b 上的脑提取物的亲和层析进一步分离了 TRIP8b 相互作用蛋白。通过质谱法鉴定了纯化制剂中的十六种蛋白质。网格蛋白和 AP2 复合体亚基似乎是主要的 TRIP8b 相互作用蛋白。我们的数据表明 TRIP8b 在受体介导的内吞作用中发挥作用。
Calcium-independent receptor of latrotoxin (CIRL) is an orphan heptahelical receptor implicated in regulation of exocytosis. To characterize molecular mechanisms of CIRL functioning, we searched for its intracellular partners using the yeast two-hybrid SR system with the cytoplasmic C-terminal fragment of CIRL as bait. One of the interacting proteins was identified as TRIP8b, a putative cytosolic adapter protein with multiple tetratricopeptide repeats. To understand functional significance of CIRL-TRIP8b interaction, we further isolated TRIP8b-interacting proteins by affinity chromatography of brain extracts on immobilized recombinant TRIP8b. Sixteen proteins were identified by mass spectrometry in the purified preparations. Clathrin and subunits of AP2 complex appeared to be the major TRIP8b-interacting proteins. Our data suggest a role of TRIP8b in receptor-mediated endocytosis.