Rotational dynamics of the Fc epsilon receptor on mast cells monitored by specific monoclonal antibodies and IgE.
The rotational motions of the type I receptor for the Fc epsilon domains (Fc epsilon RI) present on mast cells were investigated by measuring the phosphorescence emission and anisotropy decay kinetics of erythrosin (Er) covalently bound to several Fc epsilon RI-specific macromolecular ligands. The latter consisted of three murine monoclonal antibodies (IgG class) raised against the Fc epsilon RI of rat mast cells (RBL-2H3 line), their Fab fragments, and a murine monoclonal IgE. Different anisotropy decay patterns were observed for the three monovalent Er-Fab fragments bound to the Fc epsilon RI, reflecting the rotational motion of the Fe epsilon RI reported by each specific macromolecular probe bound to its particular epitope. Internal motions of the tethered Er-labeled ligands may also contribute to the observed anisotropy decay, particularly in the case of cell-bound IgE. The results corroborate an earlier study with rat Er-IgE in which the Fc epsilon RI-IgE complex was shown to be mobile throughout the temperature range examined (5-37 degrees C). The anisotropy decays of the three Er-labeled, Fc epsilon RI-specific intact mAbs bound to cells also differed markedly. Whereas the decay curves of one mAb (H10) were characterized by temperature-dependent positive amplitudes and rather short rotational correlation times, the decay of a second mAb (J17) showed complex qualitative variations with temperature, and in the case of the third antibody (F4), there was no apparent decay of anisotropy over the time and temperature ranges examined.(ABSTRACT TRUNCATED AT 250 WORDS)
Pecht
I
I
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Ortega
E
E
Jovin
T M
TM
D000911:Antibodies, Monoclonal / Q000276:immunology; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology; D003432:Cross-Linking Reagents / Q000378:metabolism; D004923:Erythrosine / Q000378:metabolism; D005454:Fluorescence Polarization; D007073:Immunoglobulin E / Q000276:immunology; D007700:Kinetics; D008024:Ligands; D008407:Mast Cells / Q000737:chemistry*; D011940:Receptor Aggregation; D011961:Receptors, Fc / Q000737:chemistry* / Q000276:immunology; D012399:Rotation; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D008024:Ligands; D011961:Receptors, Fc; D007073:Immunoglobulin E; D004923:Erythrosine