Characterization of the neisserial lipid-modified azurin bearing the H.8 epitope.
The pathogenic Neisseria have multiple genes encoding proteins that bind monoclonal antibody (MAb) H.8. We previously reported the cloning and sequencing of a meningococcal gene (laz) encoding an H.8 MAb-binding protein with a consensus lipoprotein processing site, an N-terminal domain containing the epitope for H.8 MAb binding, and a C-terminal domain with extensive similarity to the sequences of azurins from other organisms. In the current study, we showed that the product of the cloned gene could be labelled with palmitic acid, that it was subject to globomycin-sensitive processing, and that it was immunologically cross-reactive with azurin from Pseudomonas aeruginosa. All neisserial species tested, both pathogens and commensals, produced a protein recognized by anti-azurin serum. Southern blots with oligonucleotide probes specific for the azurin domain of the gene showed that it was present in a single copy in the chromosome; it was highly conserved in gonococci and meningococci, and less conserved in commensal Neisseria species.
Woods
J P
JP
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill 27599.
Dempsey
J F
JF
Kawula
T H
TH
Barritt
D S
DS
Cannon
J G
JG
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001400:Azurin / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000235:genetics*; D005798:Genes, Bacterial; D015151:Immunoblotting; D008074:Lipoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009343:Neisseria / Q000235:genetics / Q000276:immunology*; D019308:Palmitic Acid; D010169:Palmitic Acids; D021382:Protein Sorting Signals / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D000939:Epitopes; D008074:Lipoproteins; D010169:Palmitic Acids; D021382:Protein Sorting Signals; D001400:Azurin; D019308:Palmitic Acid