Purification, characterization, and toxicity of the sulfhydryl-activated hemolysin listeriolysin O from Listeria monocytogenes.
We purified and characterized an extracellular hemolysin produced by Listeria monocytogenes. Hemolysin production was greatly enhanced by growing bacteria in resin (Chelex)-treated medium. This hemolysin was separated as a homogeneous protein of 60,000 daltons by using thiol-disulfide exchange affinity chromatography. This protein was a sulfhydryl-activated toxin, termed listeriolysin O, which shared the classical properties of other bacterial sulfhydryl-activated toxins: inhibition by very low amounts of cholesterol; activation by reducing agents and suppression of the lytic activity by oxidation; antigenic cross-reactivity with streptolysin O. However, listeriolysin O differed remarkably from the other sulfhydryl-activated toxins in that its cytolytic activity towards erythrocytes from various animal species was maximum at low pH (approximately 5.5) and was undetectable at pH 7.0. This suggests that the lytic activity of the toxin in host tissues might be better expressed in the acidic microenvironment, including macrophage phagosomes where bacteria presumably replicate. Listeriolysin O was lethal to mice (50% lethal dose of ca. 0.8 microgram) and induced a rapid inflammatory reaction when injected intradermally. These results favor the view that listeriolysin O might play a major role during intracellular replication of L. monocytogenes, ultimately promoting death of infected macrophages.
D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000633:toxicity; D001427:Bacterial Toxins / Q000302:isolation & purification* / Q000633:toxicity; D002784:Cholesterol / Q000494:pharmacology; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins / Q000302:isolation & purification* / Q000633:toxicity; D006461:Hemolysis; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007249:Inflammation; D008089:Listeria monocytogenes / Q000032:analysis* / Q000472:pathogenicity; D051379:Mice; D008970:Molecular Weight; D013438:Sulfhydryl Compounds
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D013438:Sulfhydryl Compounds; D002784:Cholesterol; C038705:hlyA protein, Listeria monocytogenes