A method for purification of listeriolysin O from a hypersecretor strain of Listeria monocytogenes.
A simple and convenient method for the purification of the hemolytic toxin listeriolysin O (LLO) from Listeria monocytogenes is described. Supernatants from bacteria cultures were purified by application to a CH2 spiral cartridge concentrator (Amicon) and ion exchange chromatography. A critical step is removal of contaminating RNA. The purified proteins had characteristics described for bacterial thiol-activated hemolysins: activation by a reducing agent (DTT) and inactivation by cholesterol. In addition, the molecular weight of 58, 000 and pH-dependent hemolytic activity of this purified protein are consistent with the previously published characteristics of LLO.
Department of Medicine, Division of Gastroenterology-Hepatology, Farmington, Connecticut 06030, USA.
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CM
Walton
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Wu
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Wu
D001427:Bacterial Toxins; D002784:Cholesterol / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D017077:Culture Media, Conditioned / Q000737:chemistry; D004229:Dithiothreitol / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000302:isolation & purification*; D006460:Hemolysin Proteins; D006461:Hemolysis / Q000187:drug effects; D008089:Listeria monocytogenes / Q000378:metabolism*
D001427:Bacterial Toxins; D017077:Culture Media, Conditioned; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D002784:Cholesterol; C038705:hlyA protein, Listeria monocytogenes; D004229:Dithiothreitol