Immunochemical characterization of the formyl peptide receptor moieties on human neutrophils.
The neutrophil (PMN) receptor for formylated peptides such as N-formyl-l-methionyl-l-leucyl-l-phenylalanine (FMLP) is involved in binding and subsequent response to certain chemotactic stimuli. The receptor on human PMN has been reported to consist of several glycoprotein components, ranging in size from 43-94 kDa. Furthermore, FMLP receptors on human PMN have been shown to contain both high and low affinity states. In this study, the receptor was purified by subjecting solubilized PMN plasma membrane components to FMLP-affinity chromatography, and was found to be comprised of four components, one of 68 kDa, and the others of 94, 48, and approximately 40 kDa. Only the 68, the 94, and the approximately 40 kDa components specifically bound a radioiodinated FMLP analogue. To further characterize these components, a battery of monoclonal antibodies reactive against the FMLP receptor was prepared. Seven monoclonal antibodies were selected on the basis of their reactivity with the 68 kDa receptor component. Some of these antibodies also cross-react with the 48 kDa component, suggesting that the 68 and the 48 kDa receptor moieties are immunologically related. These antibodies reacted with normal human neutrophils, but not with lymphocytes, or unstimulated HL-60 cells. Furthermore, the presence of 20 nmol of FMLP inhibited the binding of five of the anti-receptor antibodies to whole PMN. These results suggest that the epitopes recognized by these five antibodies may possibly be involved in FMLP binding.
De Nardin
E
E
Department of Oral Biology, State University of New York, Buffalo 14214.
Genco
R J
RJ
D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D001667:Binding, Competitive; D003429:Cross Reactions; D000939:Epitopes; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007120:Immunochemistry; D051379:Mice; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism*; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000939:Epitopes; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D009240:N-Formylmethionine Leucyl-Phenylalanine