Identification of novel human WW domain-containing proteins by cloning of ligand targets
Identification of novel human WW domain-containing proteins by cloning of ligand targets
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DOI：
10.1074/jbc.272.23.14611
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发表时间：
1997-06-06
影响因子：
4.8
通讯作者：
Fowlkes, DM
中科院分区：
文献类型：
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作者：
Pirozzi, G;McConnell, SJ;Fowlkes, DM
最近描述的蛋白模块由35-40个被取代的残基组成，称为WW结构域，已经在许多不同的蛋白质中鉴定，包括肌营养不良蛋白和Yes相关蛋白（雅普）。雅普的两个推定配体，称为WBP-1和WBP-2，先前已经发现含有几个由PPPPY残基（PY基序）组成的短肽区域，其介导与雅普的WW结构域的结合。虽然WW结构域的功能仍有待阐明，但这些观察结果强烈支持WW结构域在蛋白质-蛋白质相互作用中的作用。在这里，我们报告了三个新的人类cDNA编码共9个WW域的分离，使用一种新开发的方法称为COLT（克隆的配体目标），其中模块化的蛋白质结构域的快速克隆是通过筛选cDNA表达文库与特定的肽配体。发现的两个新基因似乎是一个蛋白质家族的成员，包括Rsp 5和Nedd-4，它们具有泛素蛋白连接酶活性。此外，我们证明，肽对应的PY和PY样基序存在于几个已知的信号或调节蛋白，包括RasGAP，AP-2，p53 BP-2（p53结合蛋白-2），白细胞介素-6受体-α，氯离子通道CLCN 5，和上皮钠通道ENaC，可以选择性地结合到某些这些新的WW域。
A recently described protein module consisting of 35-40 semiconserved residues, termed the WW domain, has been identified in a number of diverse proteins in eluding dystrophin and Yes-associated protein (YAP). Two putative ligands of YAP, termed WBP-1 and WBP-2, have been found previously to contain several short peptide regions consisting of PPPPY residues (PY motif) that mediate binding to the WW domain of YAP. Although the function(s) of the WW domain remain to be elucidated, these observations strongly support a role for the WW domain in protein-protein interactions. Here we report the isolation of three novel human cDNAs encoding a total of nine WW domains, using a newly developed approach termed COLT (cloning of ligand targets), in which the rapid cloning of modular protein domains is accomplished by screening cDNA expression libraries with specific peptide ligands. Two of the new genes identified appear to be members of a family of proteins, including Rsp5 and Nedd-4, which have ubiquitin-protein ligase activity. In addition, we demonstrate that peptides corresponding to PY and PY-like motifs present in several known signaling or regulatory proteins, including RasGAP, AP-2, p53BP-2 (p53-binding protein-2), interleukin-6 receptor-alpha, chloride channel CLCN5, and epithelial sodium channel ENaC, can selectively bind to certain of these novel WW domains.