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.
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Isao
I
Watanabe
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Takamasa
T
Nomura
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Takanari
T
Tominaga
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Kazuhiro
K
Yamamoto
Department of Bacteriology, Showa University School of Medicine, Tokyo 142-8555, Japan.
Chikara
C
Kohda
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Ikuo
I
Kawamura
Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Masao
M
Mitsuyama
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