1. Epitope identification for human neutrophil flavocytochrome b monoclonals 48 and 449.
- Author
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Burritt JB, Fritel GN, Dahan I, Pick E, Roos D, and Jesaitis AJ
- Subjects
- Amino Acid Sequence, Antibodies, Monoclonal immunology, Consensus Sequence, Cytochrome b Group chemistry, Epitope Mapping, Epitopes blood, Humans, Membrane Glycoproteins chemistry, Membrane Glycoproteins immunology, Models, Molecular, Molecular Sequence Data, NADPH Dehydrogenase chemistry, NADPH Dehydrogenase immunology, NADPH Oxidase 2, Peptide Library, Phosphoproteins chemistry, Phosphoproteins immunology, Protein Conformation, Protein Subunits, Cytochrome b Group immunology, Epitopes chemistry, Membrane Transport Proteins, NADPH Oxidases, Neutrophils chemistry
- Abstract
Flavocytochrome b558 (Cyt b) is important in generating superoxide and other toxic oxygen species involved in inflammation and host defense. Monoclonal antibodies (mAbs) 48 and 449 bind the gp91Phox and p22phox subunits of Cyt b, respectively, and have been used to characterize this enzyme complex. Until now, data were unavailable to predict which regions of the protein were bound by each antibody. Random sequence phage-display peptide library analysis of each antibody was used to select peptides that mimic the sequence of each protein epitope. Phage sequences selected by mAb 48 presented the consensus peptide sequence, DRDVXTGL, which closely resembles 498EKDVITGL505 of gp91Phox. Phage selected by mAb 449 contributed the consensus WRWPGPQVL, resembling in part 182GPQV185 of p22phox. Confirmation for this second epitope was provided by peptide walking analysis. Identifying the protein residues bound by these antibodies makes each a more informative probe for Cyt b analysis.
- Published
- 2000
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