Biochemical and NMR characterization of the interactions of Vav2-SH2 domain with lipids and the EphA2 juxtamembrane region on membrane

Biochem J. 2020 Oct 16;477(19):3791-3801. doi: 10.1042/BCJ20200300.


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.

Keywords: EphA2; NMR; SH2 domain; VAV2; lipid binding; molecular interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ephrin-A2 / chemistry*
  • Ephrin-A2 / metabolism
  • Humans
  • Membranes, Artificial*
  • Phosphatidylinositol 4,5-Diphosphate / chemistry*
  • Phosphatidylinositol 4,5-Diphosphate / metabolism
  • Phosphatidylinositol Phosphates / chemistry*
  • Phosphatidylinositol Phosphates / metabolism
  • Proto-Oncogene Proteins c-vav / chemistry*
  • Proto-Oncogene Proteins c-vav / metabolism
  • Receptor, EphA2
  • src Homology Domains


  • EPHA2 protein, human
  • Ephrin-A2
  • Membranes, Artificial
  • Phosphatidylinositol 4,5-Diphosphate
  • Phosphatidylinositol Phosphates
  • Proto-Oncogene Proteins c-vav
  • VAV2 protein, human
  • phosphatidylinositol 3,4,5-triphosphate
  • Receptor, EphA2