Sudol M, Chen H I, Bougeret C, Einbond A, Bork P
Laboratory of Molecular Oncology, Rockefeller University, New York, NY 10021, USA.
FEBS Lett. 1995 Aug 1;369(1):67-71. doi: 10.1016/0014-5793(95)00550-s.
We have identified, characterized and cloned human, mouse and chicken cDNA of a novel protein that binds to the Src homology domain 3 (SH3) of the Yes proto-oncogene product. We subsequently named it YAP for Yes-associated protein. Analysis of the YAP sequence revealed a protein module that was found in various structural, regulatory and signaling molecules. Because one of the prominent features of this sequence motif is the presence of two conserved tryptophans (W), we named it the WW domain. Using a functional screen of a cDNA expression library, we have identified two putative ligands of the WW domain of YAP which we named WBP-1 and WBP-2. Peptide sequence comparison between the two partial clones revealed a homologous proline-rich region. Binding assays and site-specific mutagenesis have shown that the proline-rich motif binds with relatively high affinity and specificity to the WW domain of YAP, with a preliminary consensus that is different from the SH3-binding PXXP motif. This suggests that the WW domain has a role in mediating protein-protein interactions via proline-rich regions, similar but distinct from Src homology 3 (SH3) domains. Based on this finding, we hypothesize that additional protein modules exist and that they could be isolated using proline-rich peptides as functional probes.
我们已经鉴定、表征并克隆了一种与Yes原癌基因产物的Src同源结构域3（SH3）结合的新型蛋白质的人、小鼠和鸡的cDNA。我们随后将其命名为Yes相关蛋白YAP。对YAP序列的分析揭示了一个存在于各种结构、调节和信号分子中的蛋白质模块。由于该序列基序的一个显著特征是存在两个保守的色氨酸（W），我们将其命名为WW结构域。通过对cDNA表达文库的功能筛选，我们鉴定出了YAP的WW结构域的两个假定配体，我们将其命名为WBP-1和WBP-2。两个部分克隆之间的肽序列比较揭示了一个富含脯氨酸的同源区域。结合试验和位点特异性诱变表明，富含脯氨酸的基序以相对较高的亲和力和特异性与YAP的WW结构域结合，初步的共有序列不同于与SH3结合的PXXP基序。这表明WW结构域在通过富含脯氨酸的区域介导蛋白质-蛋白质相互作用中起作用，与Src同源结构域3（SH3）相似但不同。基于这一发现，我们推测还存在其他蛋白质模块，并且可以使用富含脯氨酸的肽作为功能探针来分离它们。
