Salcini A E, Confalonieri S, Doria M, Santolini E, Tassi E, Minenkova O, Cesareni G, Pelicci P G, Di Fiore P P
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy.
Genes Dev. 1997 Sep 1;11(17):2239-49. doi: 10.1101/gad.11.17.2239.
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. Based on the characteristics of EH-containing and EH-binding proteins, we propose that EH domains are involved in processes connected with the transport and sorting of molecules within the cell.
EH是最近在信号转导蛋白Eps15和Eps15R以及酵母线虫的其他几种蛋白质中发现的一种蛋白质-蛋白质相互作用结构域。我们发现，Eps15和Eps15R的EH结构域在体外与含有天冬酰胺-脯氨酸-苯丙氨酸（NPF）基序的肽段结合。用EH结构域直接筛选表达文库得到了许多推定的EH相互作用蛋白，所有这些蛋白都具有NPF基序，且该基序被证明是相互作用的原因。这些相互作用蛋白包括NUMB的人类同源物（果蝇的一个发育调控基因）以及HIV REV蛋白的细胞辅因子RAB。我们证明了Eps15与NUMB和RAB的共免疫沉淀。最后，含NPF肽段与细胞蛋白的体外结合以及EST数据库筛选证实了哺乳动物中存在一个含EH蛋白家族。基于含EH蛋白和与EH结合蛋白的特性，我们提出EH结构域参与细胞内分子的运输和分选相关过程。
