title: The mechanism by which proteolysis enhances the ligand-binding activity of guinea pig type II Fc receptor for IgG (FcgammaRIIB).
abstract: We have previously shown that the ligand-binding activity of type II Fc receptor for IgG (FcgammaRIIB) on guinea pig peripheral blood polymorphonuclear leukocytes is very low and dramatically increases after treatment of the cells with proteolytic enzymes. In the present study, we analyzed the mechanism of this augmentation. We found that the protease treatment failed to enhance the binding of monomeric IgG to FcgammaRIIB, increased the binding of small immune complexes (IC) prepared under antigen-excess conditions only modestly, but markedly enhanced the binding of large IC prepared under antibody-excess conditions. These results suggest that proteolysis increases the ligand-binding avidity but not the intrinsic affinity of FcgammaRIIB. Confocal laser scanning microscopy revealed that the mobility of FcgammaRIIB on the cell surface was increased after protease treatment. In addition, transfection experiments indicated that the effect of proteolysis on IC binding to CHO cells expressing guinea pig FcgammaRIIB was strongly dependent on the receptor density. Finally, we demonstrated that the transmembrane and cytoplasmic domains of FcgammaRIIB were not involved in the proteolysis-induced augmentation of IC binding. Together our results suggest that the mobility of FcgammaRIIB, which may be restricted due to the association of the ectodomain of the receptor with unknown membrane proteins, is enhanced by proteolysis, allowing the receptors to bind multivalent ligands more readily and hence with higher avidity.
authors.affiliation: Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, and Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Kita-12 Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Isashi
authors.forename: T
authors.initials: T
authors.lastname: Yamashita
authors.forename: S
authors.initials: S
authors.lastname: Nagasawa
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
authors.forename: M
authors.initials: M
authors.lastname: Murakami
authors.forename: T
authors.initials: T
authors.lastname: Uede
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000276:immunology / Q000378:metabolism*; D015703:Antigens, CD / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D016466:CHO Cells; D006224:Cricetinae; D010450:Endopeptidases / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D008024:Ligands; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D017452:Receptors, IgG / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D012995:Solubility; D013347:Subcellular Fractions / Q000276:immunology / Q000378:metabolism; D014162:Transfection
chemical_list: D000936:Antigen-Antibody Complex; D015703:Antigens, CD; C080571:Fc gamma receptor IIB; D008024:Ligands; D008565:Membrane Proteins; D017452:Receptors, IgG; D010450:Endopeptidases