Concurrent and independent binding of Fcgamma receptors IIa and IIIb to surface-bound IgG.
Fc receptor-antibody interactions are key mechanisms through which antibody effector functions are mediated. Neutrophils coexpress two low-affinity Fcgamma receptors, CD16b (FcgammaRIIIb) and CD32a (FcgammaRIIa), possessing overlapping ligand binding specificities but distinct membrane anchor and signaling capacities. Using K562 cell transfectants as a model, the kinetics of both separate and concurrent binding of CD16b and CD32a to surface-bound IgG ligands were studied. CD16b bound human IgG with 2-3 times higher affinity than did CD32a (A(c)K(a) = 4.1 and 1.6 x 10(-7) microm(4), respectively) and both FcgammaRs had similar reverse kinetic rates (k(r) = 0.5 and 0.4 s(-1), respectively). Because CD16b is expressed on neutrophils at a 4-5 times higher density than CD32a, our results suggest that CD16b plays the dominant role in binding of neutrophils to immobilized IgG. The question of possible cross-regulation of binding affinity between CD16b and CD32a was investigated using our multispecies concurrent binding model (Zhu and Williams, Biophys. J. 79:1850-1857, 2000). Because the model assumes independent binding (no cooperation among different species), the excellent agreement between the model predictions and the experimental data suggests that, when coexpressed on K562 cells, these two FcgammaRs do not interact in a manner that alters the kinetic rates of either molecule.
Williams
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George W. Woodruff School of Mechanical Engineering and Georgia Tech/Emory Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332-0363, USA.
Nagarajan
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Selvaraj
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Zhu
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C
D019138:Antibodies, Blocking / Q000378:metabolism; D001667:Binding, Competitive; D055592:Biophysical Phenomena; D001703:Biophysics; D004912:Erythrocytes / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D066298:In Vitro Techniques; D020014:K562 Cells; D007700:Kinetics; D008954:Models, Biological; D009504:Neutrophils / Q000276:immunology; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D017452:Receptors, IgG / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D019138:Antibodies, Blocking; D007074:Immunoglobulin G; D011947:Receptors, Antigen, B-Cell; D017452:Receptors, IgG