Paoluzi S, Castagnoli L, Lauro I, Salcini A E, Coda L, Fre' S, Confalonieri S, Pelicci P G, Di Fiore P P, Cesareni G
Department of Biology, Enrico Calef, University of Rome Tor Vergata, Rome 00133, USA.
EMBO J. 1998 Nov 16;17(22):6541-50. doi: 10.1093/emboj/17.22.6541.
The Eps homology (EH) domain is a recently described protein binding module that is found, in multiple or single copies, in several proteins in species as diverse as human and yeast. In this work, we have investigated the molecular details of recognition specificity mediated by this domain family by characterizing the peptide-binding preference of 11 different EH domains from mammal and yeast proteins. Ten of the eleven EH domains could bind at least some peptides containing an Asn-Pro-Phe (NPF) motif. By contrast, the first EH domain of End3p preferentially binds peptides containing an His-Thr/Ser-Phe (HT/SF) motif. Domains that have a low affinity for the majority of NPF peptides reveal some affinity for a third class of peptides that contains two consecutive amino acids with aromatic side chains (FW or WW). This is the case for the third EH domain of Eps15 and for the two N-terminal domains of YBL47c. The consensus sequences derived from the peptides selected from phage-displayed peptide libraries allows for grouping of EH domains into families that are characterized by different NPF-context preference. Finally, comparison of the primary sequence of EH domains with similar or divergent specificity identifies a residue at position +3 following a conserved tryptophan, whose chemical characteristics modulate binding preference.
Eps 同源（EH）结构域是一种最近才被描述的蛋白质结合模块，在人类和酵母等多种物种的几种蛋白质中以多个或单个拷贝形式存在。在这项研究中，我们通过表征来自哺乳动物和酵母蛋白质的 11 种不同 EH 结构域的肽结合偏好，研究了该结构域家族介导的识别特异性的分子细节。11 个 EH 结构域中的 10 个能够结合至少一些含有 Asn-Pro-Phe（NPF）基序的肽。相比之下，End3p 的第一个 EH 结构域优先结合含有 His-Thr/Ser-Phe（HT/SF）基序的肽。对大多数 NPF 肽亲和力较低的结构域对第三类含有两个连续带有芳香族侧链氨基酸（FW 或 WW）的肽表现出一定亲和力。Eps15 的第三个 EH 结构域以及 YBL47c 的两个 N 端结构域就是这种情况。从噬菌体展示肽库中筛选出的肽推导得到的共有序列能够将 EH 结构域分为不同家族，这些家族具有不同的 NPF 上下文偏好。最后，对具有相似或不同特异性的 EH 结构域的一级序列进行比较，发现在保守色氨酸之后的 +3 位存在一个残基，其化学特性调节结合偏好。
