Purification and further characterization of a haemolysin of Actinomyces pyogenes.
A haemolysin produced by Actinomyces pyogenes ATCC 8164 was purified from culture supernatant by ammonium sulphate and polyethylene glycol precipitation, ion-exchange chromatography on DEAE-Sephacel, and fast-protein-liquid-chromatography on Superose 12 prep grade. The purified haemolysin, designated as pyolysin, displayed a single band on poly-acrylamide gel electrophoresis, indicating a molecular weight of 55000. Additionally, using gel filtration, the same molecular weight was estimated. Further studies of the eluate of ion-exchange chromatography using isoelectric focusing also revealed a single protein band at pH 9.38 with haemolytic activity. A specific antiserum produced against pyolysin inhibited the haemolytic activity. The purity of the isolated protein was also determined by Western Blot analysis with antiserum obtained from a cow inoculated with culture supernatant from A. pyogenes and Peptococcus indolicus. The isolated pyolysin appeared to be heat-labile and displayed cytotoxic effects on poly-morphonuclear leucocytes and on pTK2 kidney cells.
Ding
H
H
Institut für Bakteriologie und Immunologie, Justus-Liebig-Universität, Giessen, Germany.
Lämmler
C
C
D000190:Actinomyces / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D006460:Hemolysin Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D006461:Hemolysis; D007614:Macropodidae; D008970:Molecular Weight; D009504:Neutrophils / Q000187:drug effects
D006460:Hemolysin Proteins