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Epitope identification for human neutrophil flavocytochrome b monoclonals 48 and 449.

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.
AuthorsJ B Burritt, G N Fritel, I Dahan, E Pick, D Roos, A J Jesaitis
JournalEuropean journal of haematology (Eur J Haematol) Vol. 65 Issue 6 Pg. 407-13 (Dec 2000) ISSN: 0902-4441 [Print] Denmark
PMID11168498 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Antibodies, Monoclonal
  • CYBB protein, human
  • Cytochrome b Group
  • Epitopes
  • Membrane Glycoproteins
  • Membrane Transport Proteins
  • Peptide Library
  • Phosphoproteins
  • Protein Subunits
  • cytochrome b558
  • CYBA protein, human
  • NADPH Oxidase
  • NADPH Dehydrogenase
Topics
  • Amino Acid Sequence
  • Antibodies, Monoclonal (immunology)
  • Consensus Sequence
  • Cytochrome b Group (chemistry, immunology)
  • Epitope Mapping
  • Epitopes (blood, chemistry)
  • Humans
  • Membrane Glycoproteins (chemistry, immunology)
  • Membrane Transport Proteins
  • Models, Molecular
  • Molecular Sequence Data
  • NADPH Dehydrogenase (chemistry, immunology)
  • NADPH Oxidase
  • Neutrophils (chemistry)
  • Peptide Library
  • Phosphoproteins (chemistry, immunology)
  • Protein Conformation
  • Protein Subunits

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