Crystal structure of a C-terminal fragment of growth arrest-specific protein Gas6. Receptor tyrosine kinase activation by laminin G-like domains

J Biol Chem. 2002 Nov 15;277(46):44164-70. doi: 10.1074/jbc.M207340200. Epub 2002 Sep 5.

Abstract

Receptor tyrosine kinases of the Axl family are activated by Gas6, the product of growth arrest-specific gene 6. Gas6-Axl signaling is implicated in cell survival, adhesion, and migration. The receptor-binding site of Gas6 is located within a C-terminal pair of laminin G-like (LG) domains that do not resemble any other receptor tyrosine kinase ligand. We report the crystal structure at 2.2-A resolution of a Gas6 fragment spanning both LG domains (Gas6-LG). The structure reveals a V-shaped arrangement of LG domains strengthened by an interdomain calcium-binding site. LG2 of Gas6-LG contains two unusual features: an alpha-helix cradled by one edge of the LG beta-sandwich and a conspicuous patch of surface-exposed hydrophobic residues. Mutagenesis of some residues in this patch reduces Gas6-LG binding to the extracellular domain of Axl as well as Axl activation in glioblastoma cells, identifying a component of the receptor-binding site of Gas6.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Calcium / metabolism
  • Crystallography, X-Ray
  • Culture Media, Serum-Free / pharmacology
  • DNA, Complementary / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Intercellular Signaling Peptides and Proteins*
  • Laminin / chemistry*
  • Ligands
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Structure, Tertiary
  • Proteins / chemistry*
  • Sequence Homology, Amino Acid
  • Tumor Cells, Cultured

Substances

  • Culture Media, Serum-Free
  • DNA, Complementary
  • Intercellular Signaling Peptides and Proteins
  • Laminin
  • Ligands
  • Proteins
  • growth arrest-specific protein 6
  • Calcium