Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b.
The P2 porin protein is the most abundant protein in the outer membrane of Haemophilus influenzae type b (Hib). Biochemical and immunochemical techniques were used to characterize the P2 proteins from a number of different Hib strains. P2 proteins from Hib outer membrane vesicles were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose for in situ tryptic digestion. Solid-phase tryptic digests of P2 from eight Hib strains were resolved by high-pressure liquid chromatography and shown to be similar if not identical. Radioimmunoprecipitation analysis involving Hib cells (containing intrinsically radiolabeled proteins or lipooligosaccharide) and Western blot (immunoblot) analysis were used to identify two P2-specific murine monoclonal antibodies (MAbs). These MAbs were shown to be reactive with 120 Hib strains tested in a colony blot radioimmunoassay. One of these MAbs bound to a surface-exposed P2 epitope that was antibody accessible on all Hib strains tested; the other MAb was directed against a P2 epitope that either was not exposed on the cell surface or was otherwise inaccessible to antibody.
Hansen
E J
EJ
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
Pelzel
S E
SE
Orth
K
K
Moomaw
C R
CR
Radolf
J D
JD
Slaughter
C A
CA
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D000939:Epitopes; D006193:Haemophilus influenzae / Q000145:classification / Q000276:immunology* / Q000648:ultrastructure; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D018272:Porins; D013045:Species Specificity; D014357:Trypsin / Q000494:pharmacology
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D018272:Porins; D014357:Trypsin