Bivalent ligand dissociation kinetics from receptor-bound immunoglobulin E: evidence for a time-dependent increase in ligand rebinding at the cell surface.
The bivalent ligand N,N'-bis[[epsilon-[(2,4- dinitrophenyl)amino]caproyl]-L-tyrosyl]cystine [(DCT)2-Cys] binds and cross-links anti-dinitrophenyl (DNP) immunoglobulin E (IgE)-receptor complexes on the cell surface of rat basophilic leukemia cells. The rate of dissociation of this bound ligand was monitored by using a fluorescence method under two different conditions. In one case the monovalent ligand DCT was added in large excess to prevent the dissociating ligand from rebinding to unoccupied antibody combining sites. Under these conditions, dissociation of the bivalent ligand from IgE-sensitized cells proceeds to completion with kinetics that are well described by two rate constants that are independent of the time of preincubation of the bivalent ligand with the cells. In the second case, dissociation of (DCT)2-Cys from cell-bound anti-DNP IgE was monitored in the presence of a large excess of anti-DNP IgE in solution that acts as a sink to absorb the dissociated ligand. Under these conditions, the bivalent ligand becomes more resistant to dissociation as the preincubation time of the bivalent ligand with the cells is increased. An increasing fraction of the bound ligand does not dissociate on a measurable time scale in the presence of this sink. The results indicate that cell-associated IgE-receptor complexes undergo a time-dependent change that facilitates the reformation of the cross-linked state when one end of the ligand dissociates to break up the existing cross-link. The possible physical basis and functional implications of these results are discussed.
Erickson
J W
JW
Department of Chemistry, Baker Laboratory, Cornell University, Ithaca, New York 14853.
Posner
R G
RG
Goldstein
B
B
Holowka
D
D
Baird
B
B
D000818:Animals; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007073:Immunoglobulin E / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008961:Models, Structural; D009842:Oligopeptides; D011961:Receptors, Fc / Q000378:metabolism*; D017455:Receptors, IgE; D013050:Spectrometry, Fluorescence; D013861:Thiocyanates; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008024:Ligands; D009842:Oligopeptides; D011961:Receptors, Fc; D017455:Receptors, IgE; D013861:Thiocyanates; C067808:N,N'-bis((epsilon-((2,4-dinitrophenyl)amino)caproyl)tyrosyl)cystine; D007073:Immunoglobulin E; D016650:Fluorescein-5-isothiocyanate