Abstract：
The Src Homology 2 (SH2) domain is an emerging biotechnology with applications in basic science, drug discovery, and even diagnostics. The SH2 domains rapid uptake into different areas of research is a direct result of the wealth of information generated on its biochemical, biological, and biophysical role in mammalian cell biology. Functionally, the SH2 domain binds and recognizes specific phosphotyrosine (pTyr) residues in the cell to mediate protein-protein interactions (PPIs) that govern signal transduction networks. These signal transduction networks are responsible for relaying growth and stress state signals to the cell\'s nucleus, ultimately effecting a change in cell biology. Protein engineers have been able to increase the affinity of SH2 domains for pTyr while also tailoring the domains\' specificity to unique amino acid sequences flanking the pTyr residue. In this way, it has been possible to develop unique SH2 variants for use in affinity-purification coupled to mass spectrometry (AP-MS) experiments, microscopy, or even synthetic biology. This chapter outlines methods to tailor the affinity and specificity of virtually any human SH2 domain using a combination of rational engineering and phage-display approaches.
摘要：
Src同源性2（SH2）领域是一种新兴的生物技术，在基础科学中具有应用，药物发现,甚至诊断。SH2领域快速吸收到不同的研究领域是其生化产生的大量信息的直接结果，生物，和哺乳动物细胞生物学中的生物物理作用。功能上,SH2结构域结合并识别细胞中特定的磷酸酪氨酸(pTyr)残基,以介导控制信号转导网络的蛋白质-蛋白质相互作用(PPIs).这些信号转导网络负责将生长和应激状态信号传递到细胞核，最终影响细胞生物学的变化。蛋白质工程师已经能够增加SH2结构域对pTyr的亲和力，同时还针对pTyr残基侧翼的独特氨基酸序列调整结构域特异性。这样,有可能开发独特的SH2变体，用于与质谱（AP-MS）实验耦合的亲和纯化，显微镜,甚至合成生物学。本章概述了使用合理工程和噬菌体展示方法的组合来定制几乎任何人类SH2结构域的亲和力和特异性的方法。
