2004 Fiscal Year Final Research Report Summary
Studies on the Interaction between Subunits of the Botulinum Toxin Complex and X-Ray Crystallography
| Project/Area Number |
15570103
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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 |
Structural biochemistry
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| Research Institution | Tokyo University of Agriculture |
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Principal Investigator |
OHYAMA Tohru Tokyo Univ.of Agriculture, Dept.of Food Science and Technology, Fac.of Bioindustry, Professor, 生物産業学部, 教授 (60318178)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Toshihiro Tokyo Univ.of Agriculture, Dept.of Food Science and Technology, Fac.of Bioindustry, Professor, 生物産業学部, 教授 (80175695)
NAKAGAWA Tomoyuki Tokyo Univ.of Agriculture, Dept.of Food Science and Technology, Fac.of Bioindustry, Assistant Professor, 生物産業学部, 講師 (70318179)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Botulinum toxin / Subunit structure / X-ray crystallography / Protein complex / Protein-protein interaction |
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| Research Abstract |
Botulinum toxin is produced as a 650 kDa toxin complex(TC) with subunit structure that non-covalently associated with five different protein components, 150 kDa neurotoxin(NT), nontoxic non-hemagglutinin(NTNHA) and three hemagglutinin subcomponents, in culture supernatant. In this study, we have clarified molecular ration of subunit components in the TC. Type D strain 4947(D-4947) produces a large amount of TC species (650 kDa L-TC,610 kDa and 540 kDa TC). They revealed same banding patterns on SDS-PAGE, but less number of HA-33/17 complexes in the TC. Based on titration experiments with HA-33/17 complex, we have concluded that complete subunit composition of the TC is a dodecamer composed of a single NT, a single NTNHA, two HA-70,four HA-33 and four HA-17 molecules. Accordingly, we have preliminary information on the analysis of the three-dimensional structure of botulinum TC by X-ray crystallography. Of the TC species, recombination of the components was found in 610 and 540 kDa TCs depending upon storage condition such as temperature, and was thus depressed by the addition of glycerol leading supply of stable TC for the experiments. Additionally, we have established new method to separate considerable amount of the NT from the D-4947 TC, which has been technically difficult to separate. Although we have attempted to crystallize TCs (M and L-TC) that were examined using dynamic light scattering from serotype C and D strains by 130 combinations of experimental condition, it has not been yet obtain large crystals for X-ray analysis.
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[Journal Article] Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal cells and erythrocytes2004
Author(s)
Fujinaga, Y., Inoue, K., Watarai, S., Sakaguchi, Y., Arimitsu, H., Lee, J.C., Jin, Y., Matsumura, T., Kabumoto, Y., Watanabe, T., Ohyama, T., Nishikawa, A., Oguma, K.
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Journal Title
Microbiology Vol.150
Pages: 1529-1538
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The receptor and transporter for internalization of Clostridium botulinum type C progenitor toxin into HT-29 cells2004
Author(s)
Nishikawa, A., Uotsu, N., Arimitsu, H., Lee, J.C., Miura, Y., Fujinaga, Y., Nakada, H., Watanabe, T., Ohyama, T., Sakano, Y., Oguma, K.
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Journal Title
Biochem.Biophys.Res.Commun. Vol.319
Pages: 327-333
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Characterization of Toxin Complex Produced by a Unique Strain of Clostridium botulinum type D 49472004
Author(s)
Hasegawa, K., Watanabe, T., Sato, H., Sagane, Y., Mutoh, S., Suzuki, T., Yamano, Y., Kouguchi, H., Takeshi, K., Kamaguchi, A., Fujinaga, Y., Oguma, K., Ohyama, T.
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Journal Title
Protein J. Vol.23
Pages: 371-378
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Molecular Characterization of GroES and GroEL Homologues from Clostridium botulinum2003
Author(s)
Sagane, Y., Hasegawa, K., Mutoh, S., Kouguchi, K., Suzuki, T., Sunagawa, H., Nakagawa, T., Kamaguchi, A., Okasaki, S., Nakayama, K., Watanabe, T., Oguma K., Ohyama, T.
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Journal Title
J.Protein Chem. Vol.22
Pages: 99-108
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Purification of Fully Activated Clostridium botulinum Serotype B Toxin for Treatment of Patients with Dystonia2003
Author(s)
Arimitsu, H., Inoue, K., Sakaguchi, Y., Lee, J.C., Fujinaga, Y., Watanabe, T., Ohyama, T., Hirst, R., Oguma, K.
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Journal Title
Infect.Immun. Vol.71
Pages: 1599-1603
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Complete Subunit Structure of Clostridium botulinum Type D Toxin Complex via Intermediate Assembly with Nontoxic Components2003
Author(s)
Mutoh, S., Kouguchi, H., Sagane, Y., Suzuki, T., Hasegawa, K., Watanabe, T., Ohyama, T.
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Journal Title
Biochemistry Vol.37
Pages: 10991-10997
Description
「研究成果報告書概要(欧文)」より
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[Book] Structure and function of the Clostridium botulinum serotypes C and D toxin complex2005
Author(s)
Watanabe T., Sagane, Y., Mutoh, S., Suzuki, T., Hasegawa, K., Kouguchi, H., Ohyama, T.
Total Pages
17-33
Publisher
Research Signpost
Description
「研究成果報告書概要(欧文)」より
