Extracellular mutations of non-obese diabetic mouse FcgammaRI modify surface expression and ligand binding

J Biol Chem. 1996 Jul 19;271(29):17091-9. doi: 10.1074/jbc.271.29.17091.

Abstract

The non-obese diabetic mouse (NOD) expresses a unique form of the high affinity receptor for IgG (FcgammaRI), containing multiple mutations that result in substitutions and insertions of amino acids and a truncated cytoplasmic tail. As a result of these major changes, receptor affinity for IgG increases 10-fold over that of wild-type FcgammaRI from BALB/c mice, while the specificity for ligand is retained. Kinetic analysis revealed that while the association rate of IgG with FcgammaRI from NOD mice (FcgammaRI-NOD) and FcgammaRI from BALB/c mice (FcgammaRI-BALB) is similar, IgG bound much more tightly to FcgammaRI-NOD as revealed by a profoundly diminished dissociation rate. Transfection studies demonstrated that FcgammaRI-NOD was expressed at one-tenth of the level of FcgammaRI-BALB. Although mouse FcgammaRI was previously not known to associate with the FcepsilonRI gamma-subunit, transfection of COS-7 cells demonstrates that like human FcgammaRI, mouse FcgammaRI is also able to associate with this signaling subunit. Furthermore, expression levels of FcgammaRI-NOD were not restored by the presence of the FcepsilonRI gamma-subunit. The difference in the levels of expression was mapped to mutations in the extracellular region of FcgammaRI-NOD as replacement of the extracellular domains with those of human FcgammaRI or FcgammaRI-BALB restored expression to that of human FcgammaRI or FcgammaRI-BALB.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bone Marrow / immunology
  • Bone Marrow Cells
  • Cell Line
  • Chlorocebus aethiops
  • DNA Primers
  • DNA, Complementary
  • Diabetes Mellitus, Type 1 / genetics
  • Diabetes Mellitus, Type 1 / immunology
  • Flow Cytometry
  • Humans
  • Immunoglobulin G / metabolism*
  • Kinetics
  • Ligands
  • Macrophages / immunology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred NOD
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Polymerase Chain Reaction
  • Receptors, IgG / biosynthesis
  • Receptors, IgG / genetics*
  • Receptors, IgG / metabolism*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Spleen / immunology
  • Substrate Specificity
  • Transfection

Substances

  • DNA Primers
  • DNA, Complementary
  • Immunoglobulin G
  • Ligands
  • Receptors, IgG
  • Recombinant Fusion Proteins