Purification of Pseudomonas aeruginosa exoenzyme S.
Pseudomonas aeruginosa produces two distinct ADP-ribosyl transferases, exotoxin A and exoenzyme S, which differ in a number of properties including substrate specificity. Exoenzyme S was purified from culture supernatants of P. aeruginosa DG1. The procedure for purification consists of four major steps: ammonium sulfate precipitation, anion-exchange chromatography on DEAE-Sephacel, acetone precipitation in the presence of 1 M NaCl, and G-100 Superfine gel filtration chromatography. Exoenzyme S was monitored during purification by an assay for ADP-ribosyl transferase activity, mouse toxicity, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified material exhibited ADP-ribosyl transferase activity, reacted with monoclonal antibodies to exoenzyme S, and was toxic to mice and a variety of tissue culture cell lines.
D036002:ADP Ribose Transferases; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001427:Bacterial Toxins; D005098:Exotoxins / Q000276:immunology / Q000633:toxicity; D051379:Mice; D010430:Pentosyltransferases / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000276:immunology; D013379:Substrate Specificity; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D000596:Amino Acids; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D005098:Exotoxins; D010430:Pentosyltransferases; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A; C444820:exoenzyme S