Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome
Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome
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DOI：
10.1038/sj.embor.7400596
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发表时间：
2006-02-01
期刊：
影响因子：
7.7
通讯作者：
Saksela, K
中科院分区：
文献类型：
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作者：
Kärkkäinen, S;Hiipakka, M;Saksela, K
我们已经确定人类基因组包含296个不同的Src同源性3（SH 3）结构域，并将它们克隆到噬菌体展示载体中。这提供了一个强大的和公正的系统，用于同时测定完整的人SH 3蛋白质组与目标靶蛋白的最强结合，而没有短线性肽配体或蛋白质相互作用筛选的更间接方法的混淆变量所带来的限制。涉及三种配体蛋白，人类免疫缺陷病毒-1 Nef，p21激活激酶（PAK）2和ADAM 15的研究显示，先前报道的以及新的SH 3合作伙伴与纳摩尔亲和力特异性。这表明SH 3结构域在指导细胞蛋白质相互作用方面可能比假设的更占主导地位。除了显示出潜在的重要的新的SH 3定向相互作用，这些研究还导致了新的信号蛋白的发现，如PAK 2结合衔接蛋白POSH 2和ADAM 15结合分选连接蛋白家族成员SNX 30。
We have determined the human genome to contain 296 different Src homology-3 (SH3) domains and cloned them into a phage-display vector. This provided a powerful and unbiased system for simultaneous assaying of the complete human SH3 proteome for the strongest binding to target proteins of interest, without the limitations posed by short linear peptide ligands or confounding variables of more indirect methods for protein interaction screening. Studies involving three ligand proteins, human immunodeficiency virus-1 Nef, p21-activated kinase (PAK) 2 and ADAM15, showed previously reported as well as novel SH3 partners with nanomolar affinities specific for them. This argues that SH3 domains may have a more dominant role in directing cellular protein interactions than has been assumed. Besides showing potentially important new SH3-directed interactions, these studies also led to the discovery of novel signalling proteins, such as the PAK2-binding adaptor protein POSH2 and the ADAM15-binding sorting nexin family member SNX30.