Identification of biotinylated enzymes involved in regulation of the toxin synthesis by Clostridium difficile.
| Project/Area Number |
13670265
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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 | Kanazawa University |
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Principal Investigator |
MAEGAWA Tsuneo Kanazawa University, Graduate School of Medical Science, Assistant Professor, 医学系研究科, 助手 (50283114)
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| Co-Investigator(Kenkyū-buntansha) |
KARASAWA Tadahiro Kanazawa University, Graduate School of Medical Science, Associate Professor, 医学系研究科, 助教授 (90251917)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Clostridium difficile / Biotinylated protein / FGAM synthetase / Glutamine synthesis / 2-D PAGE / 遺伝子クローニング / シャトルベクター |
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| Research Abstract |
According to this aa sequence, a ORF was selected from the genome data of C. difficile 630 (http://www.sanger.ac.uk/Projects/C_difficile/), and this gene product was revealed to be a homologue of D-proline reductase of Clostridium sticklandii.
Among intracellular proteins of Clostridium difficile KZ 1647, 13 proteins were reacted to streptavidin and 5 proteins were reacted to anti-biotin Mab when analyzed by western blotting. No proteins were reacted to both streptavidin and anti-biotin Mab.
Purification of those proteins was very difficult probably because of departure of biotin molecule from the biotinylated proteins during purification processes. However, one protein had been keeping the reactivity to streptavidin even after chromatography and 2-D PAGE, and N-terminal sequence of this protein was VEEKKLLRSLTKKHF.
D-Proline reductase produces 5-aminopentanoate from D-proline. 5-Aminopentanoate and 2-oxoglutarate are converted to 5-oxopentanoate and L-glutamate by another enzyme, suggesting that D-proline reductase is involved in the regulation of toxin synthesis by C. difficile via glutamine supply.
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Report
(3 results)
Research Products
(4 results)
