title: Human Fc receptor-like 5 binds intact IgG via mechanisms distinct from those of Fc receptors.
abstract: Fc receptor-like (FCRL) 5 regulates B cell Ag receptor signaling and has been reported to bind aggregated IgG. Using surface plasmon resonance, we analyzed the interaction of native IgG samples with FCRL5, revealing a complex binding mechanism, where isotype is just one factor. FCRL5 bound IgG1 and IgG4 with ~1 μM KD, whereas the interaction with IgG3 was a magnitude weaker. However, IgG2 samples displayed a wide range of affinities, indicating that additional factors affect binding. We used a panel of 19 anti-FCRL5 mAbs with defined reactivity to identify domains involved in ligand binding. Six mAbs blocked IgG binding, indicating critical roles of FCRL5 domains 1 and 3, as well as epitopes at the domain 1/2 and domain 2/3 boundaries. We found that only glycosylated IgG containing both Fab arms and the Fc region bound with high affinity. Furthermore, the presence of sialic acid in the IgG carbohydrate altered FCRL5 binding. The interaction of IgG and FCRL5 consisted of two kinetic components, suggesting a complex binding mechanism. We established that the IgG-Fc and IgG-F(ab')2 fragments bind FCRL5 independently but with low affinity, revealing the mechanism behind the two-step binding of whole IgG. This complex binding mechanism is distinct from that of Fc receptors, which bind through the Fc. We propose that FCRL5 is a new type of receptor that recognizes intact IgG, possibly enabling B cells to sense Ig quality. Recognition of undamaged IgG molecules by FCRL5 could allow B cells to engage recently produced Abs.
authors.affiliation: Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Franco
authors.forename: Bazarragchaa
authors.initials: B
authors.lastname: Damdinsuren
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ise
authors.forename: Jessica
authors.initials: J
authors.lastname: Dement-Brown
authors.forename: Huifang
authors.initials: H
authors.lastname: Li
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nagata
authors.forename: Mate
authors.initials: M
authors.lastname: Tolnay
mesh_terms: D000915:Antibody Affinity / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D007700:Kinetics; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D011961:Receptors, Fc / Q000378:metabolism*; D011994:Recombinant Proteins
chemical_list: D000939:Epitopes; C432367:FCRL5 protein, human; D007074:Immunoglobulin G; D011956:Receptors, Cell Surface; D011961:Receptors, Fc; D011994:Recombinant Proteins