Purification and characterization of a wall protein antigen from Clostridium botulinum type A.
A wall surface protein, designated antigen S, was extracted from Clostridium botulinum type A strain 190L with 0.1% Brij 58-2 M LiCl and purified sequentially by acetone pecipitation, ion-exchange chromatography, hydroxyapatite chromatography, chromatofocusing, and gel filtration. Crossed immunoelectrophoresis of the purified antigen S preparation against homologous multispecific antiserum to whole cells revealed only a single precipitin line. Antigen S had an apparent molecular weight of about 195,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen was composed predominantly of acidic amino acid residues, and the isoelectric point was estimated to be at pH 4.75. Tryptic digestion of antigen S destroyed antigenic activity and produced one major polypeptide fragment with a molecular weight of about 44,000. Indirect immunoferritin labeling showed that antigen S was located on the outer layer of the cell wall.
D000596:Amino Acids / Q000032:analysis; D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000032:analysis; D002473:Cell Wall / Q000276:immunology; D003014:Clostridium botulinum / Q000276:immunology*; D014357:Trypsin
D000596:Amino Acids; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001426:Bacterial Proteins; D014357:Trypsin