Association of adaptor protein TRIP8b with clathrin.
Association of adaptor protein TRIP8b with clathrin.
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DOI：
10.1111/j.1471-4159.2011.07384.x
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发表时间：
2011-09
影响因子：
4.7
通讯作者：
Petrenko AG
中科院分区：
文献类型：
--
作者：
Popova NV;Deyev IE;Petrenko AG
TRIP 8b是一种脑特异性亲水性胞质蛋白，含有三肽重复序列（TPR）。以前的研究表明，该蛋白通过其TPR结构域与Rab 8b小GT3，HCN通道和G蛋白偶联受体CIRL相互作用。我们确定网格蛋白是TRIP 8b亲和基质洗脱液的主要成分。在本研究中，通过体外结合分析，我们证明了网格蛋白和TRIP 8b之间的直接相互作用。网格蛋白结合位点位于TRIP 8b分子的N-末端（不含TPR）部分，该部分含有两个参与网格蛋白结合的短基序。在转染的HEK 293细胞中，HCN 1与TRIP 8b的共表达导致通道从细胞表面移位到TRIP 8b和网格蛋白都集中的大的细胞内斑点。这些斑点与早期内体标记物部分共定位，并与溶酶体染色试剂强烈重叠。当HCN 1与TRIP 8b的网格蛋白非结合突变体共表达时，网格蛋白不易位到HCN 1和含有TRIP 8b的斑点，表明TRIP 8b独立地与HCN和网格蛋白相互作用。我们发现TRIP 8b存在于从脑组织中纯化的网格蛋白包被的囊泡部分中。用Tris碱性缓冲液从囊泡中剥离网格蛋白外壳蛋白导致TRIP 8b的伴随释放。我们的数据表明TRIP 8b通过与膜蛋白和网格蛋白外壳成分的独立相互作用在神经元内吞作用中发挥复杂的调节功能。
TRIP8b is a brain-specific hydrophilic cytosolic protein that contains tetratricopeptide repeats (TPRs). Previous studies revealed interaction of this protein via its TPR-containing domain with Rab8b small GTPase, HCN channels and G protein-coupled receptor CIRL. We identified clathrin as a major component of eluates from the TRIP8b affinity matrix. In the present study, by in vitro binding analysis we demonstrate a direct interaction between clathrin and TRIP8b. The clathrin-binding site was localized in the N-terminal (non-TPR containing) part of the TRIP8b molecule that contains two short motifs involved in the clathrin binding. In transfected HEK293 cells, co-expression of HCN1 with TRIP8b resulted in translocation of the channels from the cell surface to large intracellular puncta where both TRIP8b and clathrin were concentrated. These puncta co-localized partially with an early endosome marker and strongly overlapped with lysosome staining reagent. When HCN1 was co-expressed with a clathrin-non-binding mutant of TRIP8b, clathrin did not translocate to HCN1 and TRIP8b-containing puncta, suggesting that TRIP8b interacts with HCN and clathrin independently. We found TRIP8b present in the fraction of clathrin-coated vesicles purified from brain tissues. Stripping the clathrin coat proteins from the vesicles with Tris alkaline buffer resulted in concomitant release of TRIP8b. Our data suggest complex regulatory functions of TRIP8b in neuronal endocytosis through independent interaction with membrane proteins and components of the clathrin coat.
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影响因子：
4.8
作者：
Chen, S;Liang, MC;Ting, AE
通讯作者：
Ting, AE
影响因子：
56.9
作者：
Dell'Angelica, EC;Klumperman, J;Bonifacino, JS
通讯作者：
Bonifacino, JS
影响因子：
2.8
作者：
Popova, N. V.;Plotnikov, A. N.;Petrenko, A. G.
通讯作者：
Petrenko, A. G.
影响因子：
4.8
作者：
Lelianova, VG;Davletov, BA;Ushkaryov, YA
通讯作者：
Ushkaryov, YA
影响因子：
64.5
作者：
KEEN, JH;WILLINGHAM, MC;PASTAN, IH
通讯作者：
PASTAN, IH