title: Effects of anchor structure and glycosylation of Fcγ receptor III on ligand binding affinity.
abstract: Isoforms of the Fcγ receptor III (FcγRIII or CD16) are cell surface receptors for the Fc portion of IgG and important regulators of humoral immune responses. Different ligand binding kinetics of FcγRIII isoforms are obtained in three dimensions by surface plasmon resonance and in two dimensions by a micropipette adhesion frequency assay. We show that the anchor structure of CD16 isoforms isolated from the cell membrane affects their binding affinities in a ligand-specific manner. Changing the receptor anchor structure from full to partial to none decreases the ligand binding affinity for human IgG1 (hIgG1) but increases it for murine IgG2a (mIgG2a). Removing N-glycosylation from the CD16 protein core by tunicamycin also increases the ligand binding affinity. Molecular dynamics simulations indicate that deglycosylation at Asn-163 of CD16 removes the steric hindrance for the CD16-hIgG1 Fc binding and thus increases the binding affinity. These results highlight an unexpected sensitivity of ligand binding to the receptor anchor structure and glycosylation and suggest their respective roles in controlling allosterically the conformation of the ligand binding pocket of CD16.
authors: Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332.
authors: Ning
authors: N
authors: Jiang
authors: Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332.
authors: Wei
authors: W
authors: Chen
authors: Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332.
authors: Prithiviraj
authors: P
authors: Jothikumar
authors: Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322.
authors: Jaina M
authors: JM
authors: Patel
authors: Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322.
authors: Rangaiah
authors: R
authors: Shashidharamurthy
authors: Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322.
authors: Periasamy
authors: P
authors: Selvaraj
authors: Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332 cheng.zhu@bme.gatech.edu.|Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332.
authors: Cheng
authors: C
authors: Zhu
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D058851:GPI-Linked Proteins / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D011485:Protein Binding; D020033:Protein Isoforms; D017452:Receptors, IgG / Q000378:metabolism* / Q000502:physiology; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance
chemical_list: C496124:FCGR3A protein, human; C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; D007074:Immunoglobulin G; D008024:Ligands; D020033:Protein Isoforms; D017452:Receptors, IgG