Biochemical and NMR characterization of the interactions of Vav2-SH2 domain with lipids and the EphA2 juxtamembrane region on membrane
Biochemical and NMR characterization of the interactions of Vav2-SH2 domain with lipids and the EphA2 juxtamembrane region on membrane
复制标题
Vav2-SH2 结构域与膜上脂质和 EphA2 近膜区域相互作用的生化和 NMR 表征
DOI：
10.1042/bcj20200300
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发表时间：
2020-10-01
影响因子：
4.1
通讯作者：
Wang, Junfeng
中科院分区：
文献类型：
--
作者：
Ge, Liang;Wu, Bo;Wang, Junfeng
Vav2是Rho家族GTP酶的一种普遍存在的鸟嘌呤核苷酸交换因子，参与调节广泛的生物学过程。它通过SH2结构域与多种酪氨酸磷酸化的细胞表面受体相互作用，包括Eph家族受体。Vav2与EphA2的相互作用在EphA2介导的肿瘤血管生成中起关键作用。在这里，我们发现Vav2-SH2结构域是一个脂类结合模块，可以弱而特异地识别PI(4，5)P2和PI(3，4，5)P3脂类。利用PI(4，5)P2和PI(3，4，5)P3与Vav2-SH2结合的头基进行核磁共振化学位移微扰实验，确定了Vav2-SH2结构域中的脂类结合位点。此外，用核磁共振技术研究了Vav2-SH2与EphA2(Y594磷酸化)的磷酸化膜旁区域(JM)的相互作用。此外，我们利用含镍脂多肽的纳米盘系统，研究了Vav2-SH2与脂膜上EphA2的磷酸化JM区域的结合，揭示了膜环境在调节这种蛋白质-蛋白质识别中的作用。
Vav2 is a ubiquitous guanine nucleotide exchange factor (GEF) for Rho family GTPases that is involved in regulating a wide range of biological processes. It interacts with several tyrosine-phosphorylated cell surface receptors, including the Eph family receptors, through its SH2 domain. The interaction of Vav2 with EphA2 is crucial for EphA2-mediated tumor angiogenesis. Here we show that Vav2-SH2 domain is a lipid-binding module that can recognize PI(4,5) P2 and PI(3,4,5)P3 lipids weakly but specifically. The specific lipid-binding site in Vav2-SH2 domain was identified by NMR chemical shift perturbation experiments using the head groups of PI(4,5)P2 and PI(3,4,5)P3, both of which bind to Vav2-SH2 with millimolar binding affinities. In addition, the interaction between Vav2-SH2 and the phosphorylated juxtamembrane region (JM) of EphA2 (Y594 phosphorylated) was investigated using NMR techniques. Furthermore, by using a nickel-lipid containing peptide-based nanodiscs system, we studied the binding of Vav2-SH2 to the phosphorylated JM region of EphA2 on lipid membrane and uncovered a role of membrane environment in modulating this protein-protein recognition.