1999 Fiscal Year Final Research Report Summary
Mechanism of membrane damage induced by Clostridium perfringens α toxin
| Project/Area Number |
09670305
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Tokushima Bunri University |
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Principal Investigator |
SAKURAI Jun Tokushima Bunri University, Faculty of Pharmaceutical Sciences, Full Professor, 薬学部, 教授 (80029800)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Keiko Tokushima Bunri University, Faculty of Pharmacentical Scoences, Research Associate, 薬学部, 助手 (90170315)
OCHI Sadayuki Tokushima Bunri University, Faculty of Pharmacentical Scoences, Research Associate, 薬学部, 助手 (80268705)
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| Project Period (FY) |
1997 – 1999
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| Keywords | Clostridium perfringens / Alpha-toxin / Phospholipase C / phospholipase D / Hemolysis / Site-directed-mutagenesis / Rho / Rho-GDI |
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| Research Abstract |
We reported that C. perfringens alpha-toxin activates endogenous phospholipase C (PLC) and phospholipase D (PLD) in rabbit erythrocytes, and that the toxin activates PLC through GTP-binding protein (G-protein). However, little is known about the activation of PLD in rabbit erythrocytes treated with alpha-toxin. Recently, it has been reported that PLDs in various mammalian cells are activated through low molecular G-proteins by treatment with hormones and growth factors. Rho and Rho-GDI genes were constructed from cDNA through m-RNA in rabbit reticulocytes by PCR amplification and these genes were ligated into pGEX-5X vector. Expression and purification of these proteins were performed using E. coli JM 109 transformants carrying Rho and Rho-GDI genes. Rho-GDI inhibited the toxin-induced hemolysis and PLD activation, However, Rho did not stimulate them. Rho isolated from cytosol is reported to be a inactive form and the mutant Rho in which is replaced Gly-14 with Val to be a active form. The mutant Rho (G14V) replaced the residue by site-directed mutagenesis stimulated the toxin-induced hemolysis and PLD activation. These observation suggested that PLD activation through Rho activated by the toxin plays a important role in hemolysis of rabbit erythrocytes.
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Research Products
(14 results)
