Surface peptide mapping of protein I and protein III of four strains of Neisseria gonorrhoeae.
Whole cells and isolated outer membranes (OMs) of four strains of gonococci were surface radioiodinated with either lactoperoxidase or Iodogen (Pierce Chemical Co., Rockford, Ill.). These preparations were solubilized in sodium dodecyl sulfate and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Surface-radioiodinated protein I (PI) and PIII bands were excised from the sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and digested with alpha-chymotrypsin, and the resultant 125I-peptide fragments were resolved by high-voltage electrophoresis and thin-layer chromatography (i.e., surface peptide mapping). Radioemitting peptidic fragments were visualized by autoradiography. Results demonstrated that the PI molecule of each gonococcal strain studied had unique iodinatable peptides exposed on the surface of whole cells and OMs, whereas PIIIs appeared to have the same portion of the molecule exposed on the surface of bacteria or OMs, regardless of the gonococcal strain from which they were isolated. Many more radiolabeled peptides were seen in surface peptide maps of PIs from radiolabeled OMs than in those from radioiodinated whole cells, whereas different peptidic fragments were seen in the surface peptide maps of PIIIs from radiolabeled OMs than were seen in those from radiolabeled whole cells. These data suggest that PI may contribute strain-specific antigenic determinants and PIII may contribute cross-reactive determinants and that the surface exposure of PI and PIII is different in isolated OMs than in the OM of intact gonococci.
D001425:Bacterial Outer Membrane Proteins; D001431:Bacteriological Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D007457:Iodine Radioisotopes; D007784:Lactoperoxidase / Q000494:pharmacology; D008565:Membrane Proteins / Q000032:analysis* / Q000145:classification / Q000302:isolation & purification; D009344:Neisseria gonorrhoeae / Q000032:analysis*; D010455:Peptides / Q000032:analysis*; D014508:Urea / Q000031:analogs & derivatives / Q000494:pharmacology
D001425:Bacterial Outer Membrane Proteins; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D010455:Peptides; C012385:1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril; D014508:Urea; D007784:Lactoperoxidase