A new assay system for detection of polymeric immunoglobulin A-polymeric immunoglobulin receptor binding.
A new system for detection of polymeric immunoglobulin A (pIgA)-polymeric immunoglobulin receptor (pIgR) binding was established. Cell lysates of a mouse pIgR cDNA transfectant, 2S9.1, were incubated with mouse polymeric immunoglobulin A (pIgA) or immunoglobulin G (IgG). The resulting immunocomplexes were precipitated with protein G-Sepharose and blotted with polyclonal anti-mouse pIgR antibody. The mouse pIgR molecule was specifically precipitated with pIgA, indicating successful detection of the pIgA-pIgR complex. Using this system, the role of N-glycosylation in pIgA-pIgR binding was examined. The pIgR molecule (molecular mass 100 kDa) after complete deglycosylation by tunicamycin treatment was still able to bind to pIgA, indicating that N-glycosylation of pIgR is not necessary for pIgA-pIgR binding. This novel system will be useful for detecting pIgA-pIgR complexes containing intact pIgR molecules.
Tajima
M
M
Department of Pathology, Nihon University School of Dentistry, Tokyo, Japan.
D000818:Animals; D015153:Blotting, Western; D006031:Glycosylation / Q000187:drug effects; D007118:Immunoassay; D007071:Immunoglobulin A, Secretory / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D019056:Receptors, Polymeric Immunoglobulin / Q000378:metabolism*; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D007071:Immunoglobulin A, Secretory; D019056:Receptors, Polymeric Immunoglobulin; D014415:Tunicamycin