State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan Center for Magnetic Resonance, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, 430071, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Huaihai Institute of Technology, Lianyungang, 222005, China.
Arch Biochem Biophys. 2018 Oct 15;656:31-37. doi: 10.1016/j.abb.2018.08.012. Epub 2018 Aug 27.
SH2 domain-containing inositol 5-phosphatase 2 (SHIP2) binds with the Y1356-phosphorylated hepatocyte growth factor (HGF) receptor, c-MET, through its SH2 domain, which is essential for the role of SHIP2 in HGF-induced cell scattering and cell spreading. Previously, the experimental structure of the SH2 domain from SHIP2 (SHIP2-SH2) had not been reported, and its interaction with the Y1356-phosphorylated c-MET had not been investigated from a structural point of view. In this study, the solution structure of SHIP2-SH2 was determined by NMR spectroscopy, where it was found to adopt a typical SH2-domain fold that contains a positively-charged pocket for binding to phosphotyrosine (pY). The interaction between SHIP2-SH2 and a pY-containing peptide from c-MET (Y1356 phosphorylated) was investigated through NMR titrations. The results showed that the binding affinity of SHIP2-SH2 with the phosphopeptide is at low micromolar level, and the binding interface consists of the positively-charged pocket and its surrounding regions. Furthermore, R28, S49 and R70 were identified as key residues for the binding and may directly interact with the pY. Taken together, these findings provide structural insights into the binding of SHIP2-SH2 with the Y1356-phosphorylated c-MET, and lay a foundation for further studies of the interactions between SHIP2-SH2 and its various binding partners.
含 SH2 结构域的肌醇 5-磷酸酶 2（SHIP2）通过其 SH2 结构域与 Y1356 磷酸化的肝细胞生长因子（HGF）受体 c-MET 结合，这对于 SHIP2 在 HGF 诱导的细胞散射和细胞铺展中的作用至关重要。先前，尚未报道来自 SHIP2 的 SH2 结构域（SHIP2-SH2）的实验结构，并且尚未从结构角度研究其与 Y1356 磷酸化的 c-MET 的相互作用。在这项研究中，通过 NMR 光谱学确定了 SHIP2-SH2 的溶液结构，发现其采用典型的 SH2 结构域折叠，其中包含用于结合磷酸酪氨酸（pY）的正电荷口袋。通过 NMR 滴定研究了 SHIP2-SH2 与来自 c-MET 的含 pY 肽（Y1356 磷酸化）之间的相互作用。结果表明，SHIP2-SH2 与磷酸肽的结合亲和力处于低微摩尔水平，并且结合界面由正电荷口袋及其周围区域组成。此外，确定 R28、S49 和 R70 是结合的关键残基，并且可能直接与 pY 相互作用。综上所述，这些发现为 SHIP2-SH2 与 Y1356 磷酸化的 c-MET 结合提供了结构见解，并为进一步研究 SHIP2-SH2 与其各种结合伴侣之间的相互作用奠定了基础。
