Interactions of the mast cell function-associated antigen with the type I Fcepsilon receptor.
Clustering the mast cell function-associated antigen (MAFA), a membrane glycoprotein expressed on 2H3 cells, by its specific monoclonal antibody G63 substantially inhibits secretion normally triggered by aggregating these cells' Type I Fcepsilon receptor (FcepsilonRI). To explore possible MAFA-FcepsilonRI interactions giving rise to this inhibition, we have studied by time-resolved phosphorescence anisotropy the rotational behavior of both MAFA and FcepsilonRI as ligated by various reagents involved in FcepsilonRI-induced degranulation and MAFA-mediated inhibition thereof. From 4 to 37 degrees C the rotational correlation times (mean+/-S.D.) of FcepsilonRI-bound, erythrosin-conjugated IgE resemble those observed for MAFA-bound erythrosin-conjugated G63 Fab, 82+/-17 micros and 79+/-31 micros at 4 degrees C, respectively. Clustering the FcepsilonRI-IgE complex by antigen or by anti-IgE increases the phosphorescence anisotropy of G63 Fab and slows its rotational relaxation. Lateral diffusion of G63 Fab is also slowed by antigen clustering of the receptor. Taken together, these results suggest that unperturbed MAFA associates with clustered FcepsilonRI. They are also consistent with its interaction with the isolated receptor, a situation also suggested by FRET measurements on the system.
Song
Jinming
J
Department of Physiology, Colorado State University, Fort Collins, CO 80523, USA.
Hagen
Guy
G
Smith
Steven M L
SM
Roess
Deborah A
DA
Pecht
Israel
I
Barisas
B George
BG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D015550:Cell Degranulation; D004058:Diffusion; D005454:Fluorescence Polarization; D031541:Fluorescence Resonance Energy Transfer; D007073:Immunoglobulin E / Q000378:metabolism; D037181:Lectins, C-Type / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008407:Mast Cells / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D021962:Membrane Microdomains / Q000737:chemistry; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism; D017455:Receptors, IgE / Q000378:metabolism*; D012399:Rotation; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; C094696:Klrg1 protein, rat; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D017455:Receptors, IgE; D007073:Immunoglobulin E