Antigenic and immunogenic properties of cyanogen bromide peptides from a serotype 5 gonococcal outer membrane protein I.
Treatment of a gonococcal major outer membrane protein IB (serotype 5) with cyanogen bromide (CNBr) resulted in cleavage of PIB into three major fragments of apparent molecular weight of 15, 13, and 8 kD. The location of these peptides in the intact protein was determined by analysis of partial cleavage products. The 8 kD peptide (CB2) was found to be located in the central region of the protein. Chymotrypsin cleavage of PIB revealed a cleavage site near one of the CNBr cleavage sites. Trypsin was found to cleave the protein, either in outer membranes (OMC) or in detergent micelles, in the central CB2 fragment. These result suggest that CB2 is a part of the surface exposed region of PIB. Immunization of mice with purified PIB (serotype 5) induced antibodies against all three CB-peptides. Absorption of the sera with homologous intact OMC resulted in a complete removal of antibodies against CB2, supplying further evidence for its surface exposed nature. Antibodies against the 13 kD peptide (CB1) could not be absorbed with intact OMC, suggesting that this peptide is buried within the outer membrane. Antisera raised against CB2 of serotype 5 demonstrated a considerable cross-reactivity with heterologous outer membranes. On the contrary, intact OMC induced mainly type-specific antibodies. These data demonstrate the presence of conserved epitopes on the surface exposed CB2 peptide. These conserved epitopes are generally not very immunogenic when present in intact OMC.
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000276:immunology*; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D051379:Mice; D009344:Neisseria gonorrhoeae / Q000145:classification / Q000276:immunology*; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology*; D018272:Porins
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D010446:Peptide Fragments; D018272:Porins; C057699:porin protein, Neisseria; D003488:Cyanogen Bromide