Dependence of the lethal effect of pore-forming haemolysins of Gram-positive bacteria on cytolytic activity.
Among bacterial haemolysins, cholesterol-dependent cytolysins (CDCs) produced by various Gram-positive bacteria are known to exhibit a lethal activity in mice. In this study, recombinant CDCs of streptolysin O, pneumolysin, ivanolysin O, listeriolysin O and several listeriolysin O mutants were constructed and the relationship between cytolytic activity and the lethal activity of each recombinant protein in mice was examined. Specific activity for cytolysis was determined by a quantitative haemolytic assay. Each protein was injected intravenously into mice and the lethal activity was evaluated by measuring the time until death of the mice. The four full-length CDC proteins exhibited lethal activity and their activities were highly proportional to their cytolytic activities. Inhibition of haemolytic activity resulted in the loss of lethal activity and non-haemolytic mutants of listeriolysin O did not exhibit any lethal activity. These data clearly indicate that the lethal effect of CDC proteins is dependent on the cytolytic activity.
Watanabe
Isao
I
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Nomura
Takamasa
T
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Tominaga
Takanari
T
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Yamamoto
Kazuhiro
K
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Kohda
Chikara
C
Department of Bacteriology, Showa University School of Medicine, Tokyo 142-8555, Japan.
Kawamura
Ikuo
I
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Mitsuyama
Masao
M
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000633:toxicity; D001427:Bacterial Toxins / Q000235:genetics / Q000633:toxicity; D004252:DNA Mutational Analysis; D004195:Disease Models, Animal; D004912:Erythrocytes; D005260:Female; D006094:Gram-Positive Bacteria / Q000472:pathogenicity*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000633:toxicity; D006460:Hemolysin Proteins / Q000235:genetics / Q000633:toxicity*; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000633:toxicity; D012756:Sheep; D013301:Streptolysins / Q000235:genetics / Q000633:toxicity; D016019:Survival Analysis; D037521:Virulence Factors / Q000235:genetics / Q000633:toxicity
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins; D013301:Streptolysins; D037521:Virulence Factors; C032290:plY protein, Streptococcus pneumoniae; C025964:streptolysin O; C038705:hlyA protein, Listeria monocytogenes