Liau Nicholas P D, Laktyushin Artem, Babon Jeffrey J
The Walter and Eliza Hall Institute of Medical Research, 1G Royal Pde, Parkville, Melbourne, 3052, VIC, Australia.
Department of Medical Biology, University of Melbourne, Royal Pde, Melbourne, 3050, VIC, Australia.
Methods Mol Biol. 2017;1555:173-182. doi: 10.1007/978-1-4939-6762-9_10.
Src Homology 2 (SH2) domains are protein domains which have a high binding affinity for specific amino acid sequences containing a phosphorylated tyrosine residue. The Suppressors of Cytokine Signaling (SOCS) proteins use an SH2 domain to bind to components of certain cytokine signaling pathways to downregulate the signaling cascade. The recombinantly produced SH2 domains of various SOCS proteins have been used to undertake structural and functional studies elucidating the method of how such targeting occurs. Here, we describe the protocol for the recombinant production and purification of SOCS SH2 domains, with an emphasis on SOCS3.
Src同源2(SH2)结构域是对含有磷酸化酪氨酸残基的特定氨基酸序列具有高结合亲和力的蛋白质结构域。细胞因子信号转导抑制因子(SOCS)蛋白利用一个SH2结构域与某些细胞因子信号通路的组分结合,从而下调信号级联反应。各种SOCS蛋白的重组产生的SH2结构域已被用于进行结构和功能研究,以阐明这种靶向作用是如何发生的。在此,我们描述了SOCS SH2结构域的重组生产和纯化方案,重点是SOCS3。