Structural insight into the specific interaction between murine SHPS-1/SIRP alpha and its ligand CD47

J Mol Biol. 2008 Jan 18;375(3):650-60. doi: 10.1016/j.jmb.2007.10.085. Epub 2007 Nov 7.

Abstract

SRC homology 2 domain-containing protein tyrosine phosphatase substrate 1 (SHPS-1 or SIRP alpha/BIT) is an immunoglobulin (Ig) superfamily transmembrane receptor and a member of the signal regulatory protein (SIRP) family involved in cell-cell interaction. SHPS-1 binds to its ligand CD47 to relay an inhibitory signal for cellular responses, whereas SIRPbeta, an activating member of the same family, does not bind to CD47 despite sharing a highly homologous ligand-binding domain with SHPS-1. To address the molecular basis for specific CD47 recognition by SHPS-1, we present the crystal structure of the ligand-binding domain of murine SHPS-1 (mSHPS-1). Folding topology revealed that mSHPS-1 adopts an I2-set Ig fold, but its overall structure resembles IgV domains of antigen receptors, although it has an extended loop structure (C'E loop), which forms a dimer interface in the crystal. Site-directed mutagenesis studies of mSHPS-1 identified critical residues for CD47 binding including sites in the C'E loop and regions corresponding to complementarity-determining regions of antigen receptors. The structural and functional features of mSHPS-1 are consistent with the human SHPS-1 structure except that human SHPS-1 has an additional beta-strand D. These results suggest that the variable complementarity-determining region-like loop structures in the binding surface of SHPS-1 are generally required for ligand recognition in a manner similar to that of antigen receptors, which may explain the diverse ligand-binding specificities of SIRP family receptors.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / metabolism
  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Binding Sites
  • CD47 Antigen / genetics
  • CD47 Antigen / metabolism*
  • CHO Cells
  • Cell Adhesion
  • Cricetinae
  • Cricetulus
  • Crystallography, X-Ray
  • Dimerization
  • Disulfides / chemistry
  • Escherichia coli / genetics
  • Genetic Vectors
  • Hydrogen Bonding
  • Ligands
  • Lysine / metabolism
  • Mice
  • Models, Chemical
  • Models, Molecular
  • Molecular Sequence Data
  • Molecular Weight
  • Moloney murine leukemia virus / physiology
  • Phenylalanine / metabolism
  • Protein Binding
  • Protein Folding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Receptors, Antigen, T-Cell / chemistry
  • Receptors, Antigen, T-Cell / metabolism
  • Receptors, Immunologic / chemistry
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Retroviridae / genetics
  • Sequence Homology, Amino Acid
  • Surface Plasmon Resonance
  • Transfection

Substances

  • CD47 Antigen
  • Disulfides
  • Ligands
  • Ptpns1 protein, mouse
  • Receptors, Antigen, T-Cell
  • Receptors, Immunologic
  • Recombinant Fusion Proteins
  • SHPS-1-Fc fusion protein
  • Phenylalanine
  • Lysine
  • Alanine

Associated data

  • PDB/2YZ1