A repertoire library that allows the selection of synthetic SH2s with altered binding specificities
A repertoire library that allows the selection of synthetic SH2s with altered binding specificities
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DOI：
10.1038/sj.onc.1204654
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发表时间：
2001-08-23
期刊：
影响因子：
8
通讯作者：
Di Fiore, PP
中科院分区：
文献类型：
--
作者：
Malabarba, MG;Milia, E;Di Fiore, PP
酪氨酸磷酸化是细胞外信号传递的主要机制之一。旨在干扰这一过程的策略可能允许控制几种细胞表型。Sh2结构域通过识别磷酸酪氨酸(Py)残基来调节蛋白质-蛋白质之间的相互作用。我们通过随机突变PLC Gamma C-末端SH2结构域特异性决定区中的五个关键氨基酸位置，创建了一个SH2支架资料库。使用生物素化的磷酸肽从文库中选择合成的SH2结构域；来自天然PLC伽马-SH2配体以及不相关的SH2配体。分离的SH2与所选择的多肽具有较高的结合亲和力常数，并能与相应的蛋白质相互作用。
Tyrosine phosphorylation is one of the major mechanisms involved in the intracellular propagation of external signals. Strategies aimed at interfering with this process might allow the control of several cellular phenotypes. SH2 domains mediate protein-protein interactions by recognizing phosphotyrosine (pY) residues in the context of specific phosphopeptides. We created an SH2-scaffolded repertoire library by randomly mutagenizing five critical amino acid positions in the specificity-determining region of the PLC gamma C-terminal SH2 domain. Synthetic SH2 domains were selected from the library using biotinylated phosphopeptides; derived from a natural PLC gamma -SH2 ligand as well as unrelated SH2 ligands. The isolated SH2s displayed high binding affinity constants for the selecting peptides and were capable of interacting with the corresponding proteins.