Demonstration and isolation of phallolysin, a haemolytic toxin from Amanita phalloides.
Phallolysin, a high-molecular weight, thermo- and alcohol-labile toxin with haemolytic activity, was demonstrated in Amanita phalloides. Phallolysin was isolated from cold aqueous toadstool extracts by precipitation with 40% ammonium sulphate, ion-exchange chromatography on DEAE cellulose, gel chromatography on Biogel P-30 and electrofocussing in Ampholine. Electrofocussing in a flat pH gradient resulted in the separation of the gelchromatographically homogeneous phallolysin into phallolysin A (I.P. 8.06) and phallolysin B (I.P. 7.49). Phallolysin A exhibited 70% purity with respect to its behaviour on polyacrylamide gel electrophoresis when stained with Coomassie Brilliant Blue. On the average 0.62 mug, at least 0.20 mug of protein was equivalent to 1 haemolytic unit (HU). The i.p. LD50 for mice was found to be 1060 (930--1210) HU/kg.
D000363:Agaricales / Q000032:analysis*; D000545:Amanita / Q000032:analysis*; D000546:Amanitins / Q000032:analysis / Q000302:isolation & purification* / Q000633:toxicity; D000645:Ammonium Sulfate; D000818:Animals; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005656:Fungal Proteins / Q000032:analysis; D006460:Hemolysin Proteins / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D007928:Lethal Dose 50; D051379:Mice; D008815:Mice, Inbred Strains
D000546:Amanitins; D005656:Fungal Proteins; D006460:Hemolysin Proteins; C008824:phallolysine; D000645:Ammonium Sulfate