Binding specificity and in vivo targets of the EH domain, a novel protein-protein interaction module
Binding specificity and in vivo targets of the EH domain, a novel protein-protein interaction module
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DOI：
10.1101/gad.11.17.2239
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发表时间：
1997-09-01
影响因子：
10.5
通讯作者：
DiFiore, PP
中科院分区：
文献类型：
--
作者：
Salcini, AE;Confalonieri, S;DiFiore, PP
EH是最近发现的一个蛋白-蛋白相互作用结构域，存在于酵母线虫的信号转导Eps15和Eps15R以及其他几种蛋白中。我们发现Eps15和Eps15R的EH结构域在体外与含有天冬氨酸-脯氨酸-苯丙氨酸（NPF）基序的肽结合。直接筛选具有EH结构域的表达文库产生了许多假定的EH相互作用体，所有这些相互作用体都具有NPF基序，这些基序被证明是相互作用的原因。在这些相互作用物中，有人类同源的果蝇发育调节基因NUMB和HIV REV蛋白的细胞辅助因子RAB。我们证实Eps15与NUMB和RAB共免疫沉淀。最后，通过体外将含有npf的肽与细胞蛋白结合并筛选EST数据库，确定了哺乳动物中含有eh的蛋白家族的存在。基于含EH蛋白和ed结合蛋白的特性，我们认为EH结构域参与了细胞内分子的转运和分选过程。
EH is a recently identified protein-protein interaction domain found in the signal transducers Eps15 and Eps15R and several other proteins of yeast nematode. We show that EH domains from Eps15 and Eps15R bind in vitro to peptides containing an asparagine-proline-phenylalanine (NPF) motif. Direct screening of expression libraries with EH domains yielded a number of putative EH interactors, all of which possessed NPF motifs that were shown to be responsible for the interaction. Among these interactors were the human homolog of NUMB, a developmentally reguated gene of Drosophila, and RAB, the cellular cofactor of the HIV REV protein. We demonstrated coimmunoprecipitation of Eps15 with NUMB and RAB. Finally, in vitro binding of NPF-containing peptides to cellular proteins and EST database screening established the existence of a family of EH-containing proteins in mammals. Eased on the characteristics of EH-containing and ED-binding proteins, we propose that EH domains are involved in processes connected with the transport and sorting of molecules within the cell.