Gunn Natalie J, Gorman Michael A, Dobson Renwick C J, Parker Michael W, Mulhern Terrence D
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, 30 Flemington Road, Parkville, Victoria 3010, Australia.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Mar 1;67(Pt 3):336-9. doi: 10.1107/S1744309110053728. Epub 2011 Feb 23.
The C-terminal Src kinase (Csk) and Csk-homologous kinase (CHK) are endogenous inhibitors of the proto-oncogenic Src family of protein tyrosine kinases (SFKs). Phosphotyrosyl peptide binding to their Src-homology 2 (SH2) domains activates Csk and CHK, enhancing their ability to suppress SFK signalling; however, the detailed mechanistic basis of this activation event is unclear. The CHK SH2 was expressed in Escherichia coli and the purified protein was characterized as monomeric by synchrotron small-angle X-ray scattering in-line with size-exclusion chromatography. The CHK SH2 crystallized in 0.2 M sodium bromide, 0.1 M bis-Tris propane pH 6.5 and 20% polyethylene glycol 3350 and the best crystals diffracted to ∼1.6 Å resolution. The crystals belonged to space group P2, with unit-cell parameters a=25.8, b=34.6, c=63.2 Å, β=99.4°.
C 末端 Src 激酶（Csk）和 Csk 同源激酶（CHK）是原癌基因 Src 家族蛋白酪氨酸激酶（SFK）的内源性抑制剂。磷酸酪氨酸肽与它们的 Src 同源 2（SH2）结构域结合会激活 Csk 和 CHK，增强它们抑制 SFK 信号传导的能力；然而，这种激活事件的详细机制基础尚不清楚。CHK SH2 在大肠杆菌中表达，通过同步加速器小角 X 射线散射结合尺寸排阻色谱法将纯化的蛋白表征为单体。CHK SH2 在 0.2 M 溴化钠、0.1 M 双三羟甲基氨基甲烷 pH 6.5 和 20%聚乙二醇 3350 中结晶，最佳晶体的衍射分辨率约为 1.6 Å。晶体属于空间群 P2，晶胞参数 a = 25.8、b = 34.6、c = 63.2 Å，β = 99.4°。
