2007 Fiscal Year Final Research Report Summary
Molecular basis for cell surface architecture and evolution of bacterial flagellin by structure and unction analysis of the protein
| Project/Area Number |
17380053
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Kyoto University |
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Principal Investigator |
MURATA Kousaku Kyoto University, Grad. Schl. Agric, Professor (90142299)
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| Co-Investigator(Kenkyū-buntansha) |
MIKAMI Bunzo Kyoto Univ., Grad. Schl Agric, Professor (40135611)
HASHIMOTO Wataru Kyoto Univ., Grad. Schl. Agric, Associate Professor (30273519)
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| Project Period (FY) |
2005 – 2007
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| Keywords | Sphingomonas / Alginate / Flagellin / Flagellum / Receptor / X-ray crystallography / Pit / T4 phage |
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| Research Abstract |
Alginate-assimilating Sphingomonas sp. strain Al forms a mouth-like pit through the rearrangement of cell surface pleat molecules and incorporates a macromolecule alginate through the pit The pit functions as a concentrator for extracellular polysaccharides. Two proteins, p5 (40 kDa) and p6 (31 kDa), are inducibly expressed on the cell surface in the presence of alginate, suggesting that they are responsible for the formation of the pit In this study, the structure and function of p5 and p6 have been analyzed.
Both of p5 and p6 are similar to bacterial flagellins, although strain Al forms no flagella. A p6-disruptant shows significant growth retardation in the alginate medium, and double disruption of p5 and p6 results in growth failure, indicating that these flagellin homologs might be essential for cell viability. The cell surface of the p6-disruptant differs from that of wild-strain Al in that the formation of the pit is incomplete and the cell surface structure changes from a pleat
… More
structure to a network one. Surface plasmon resonance biosensor analysis indicated that both proteins specifically bind alginate with high affinity (dissociation constant Kd〓〜 nM). Based on these results, p5 and p6 localized on the cell surface can be identified as a receptor for external alginate.
p5 consists of N-/C-terminal α (α1 + α2) domain and central B domain. Since p5 mutant lacking α_1 domain showed no alginate-binding ability, α_1 domain is essential to bind to alginate. In comparison of p5 with the Salmonella flagellated flagellin, the structure of α (α1 + α2) domain is similar each other, indicating that the alginate-binding ability commonly observed in bacterial flagellins is due to a domain conserved. On the other hand, p5 β domain is structurally similar to the finger domain of T4 phage protein gp11 located in the hinge region between the base plate and tail fiber. The binding of the finger domain to the base plate protein gp10 is important for the attachment of the tail fiber on the bacterial cell surface. The finger domain shows an affinity with the other protein, suggesting that p5 β domain functions as an anchor interacting with the strain Al cell surface molecules and contributes to the orientation of alginate-binding a domain to the external milieu. Less
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[Journal Article] Crystal structure of a novel bacterial cell-surface flagellin binding to polysaccharide2008
Author(s)
Journal, Publications, Yukie, Maruyama, Misato, Momma, Bunzo, Mikami, Wataru, Hashimoto, Kousaku, Murata
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Journal Title
Biochemstry 47
Pages: 1393-1402
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Sequence and analysis of the 46.6-kb plasmid pAl from Sphingomonas sp. Al that corresponds to the typical IncP-113 plasmid backbone without any accessory gene2006
Author(s)
Karen, Mine, Harada, Yuji, Aso, Wataru, Hashimoto, Bunzo, Mikami, Kousaku, Murata
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Journal Title
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「研究成果報告書概要(欧文)」より
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[Journal Article] Engineered membrane superchannel improves bioremediation potential of dioxin-degrading bacteria2006
Author(s)
Yuji, Aso, Yukiko, Miyamoto, Karen, Mine Harada, Keiko, Momma, Shigeyuki, Kawai, Wataru, Hashimoto, Bunzo, Mikami, Kousaku, Murata
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Journal Title
Nature Biotechnot 24
Pages: 188-189
Description
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[Journal Article] Crystallization and preliminary X-ray analysis of alginate lyases Al-II and Al-II' from Sphingomonas sp. Al2005
Author(s)
Masayuki, Yamasaki, Kohei, Ogura, Satoko, Moriwaki, Wataru, Hashimoto, Kousaku, Murata, Bunzo, Mikami
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Journal Title
Acta Crystallographica section 61
Pages: 288-290
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Proteomics-based identification of outer-membrane proteins responsible for import of macromolecules in Sphingomonas sp. Al : alginate-binding flagellin on the cell surface2005
Author(s)
Wataru, Hashimoto, Jinshan, He, Yushin, Wada, Hirokazu, Nankai, Bunzo, Mikami, Kousaku, Murata
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Journal Title
Biochemistry 44
Pages: 13783-13794
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Structure and function of bacterial super-biosystem responsible for import and depolymerization of macromolecules2005
Author(s)
Wataru, Hashimoto, Keiko, Mamma, Yukie, Maruyama, Masayuki, Yamasaki, Bunzo, Mikami, Kousaku, Murata
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Journal Title
Bioscience, Biotechnology and Biochemistry 69
Pages: 673-692
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Al flagellin based on the crystal structure and Kousaku Murata. Polysaccharide-binding ability and cell surface localization of Sphingomonas sp2007
Author(s)
Yukie, Maruyama, Misato, Momma, Asako, Chuma, Bunzo, Mikami, Wataru, Hashimoto
Organizer
Regular Meeting of Japan Society for Bioscience, Biotechnology, and, Agrochemistry, Kansai Branch
Place of Presentation
Kyoto Prefectural University
Year and Date
2007-05-26
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Structural factors for substrate specificity of alginate lyases belonging to polysaccharide Ivase families 5 and 72007
Author(s)
Kohei, Ogura, Masayuki, Yamasaki, Wataru, Hashimoto, Bunzo, Mikami, Kousaku, Murata
Organizer
Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry
Place of Presentation
Tokyo University of Agriculture
Year and Date
2007-03-26
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Crystallographic study on structure and function relationship of flagellin on the cell surface of Sphingomonas sp. Al2007
Author(s)
Yukie, Maruyama, Misato, Momma, Bunzo, Mikami, Wataru, Hashimoto, Kousaku, Murata
Organizer
Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry
Place of Presentation
Tokyo University of Agriculture
Year and Date
2007-03-25
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Catalytic reaction mechanism and substrate specificity of PL7 alginate lyase2006
Author(s)
Kohei, Ogura, Masayuki, Yamasaki, Wataru, Hashimoto, Bunzo, Mikami, Kousaku, Murata
Organizer
Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry
Place of Presentation
Kyoto Women's University
Year and Date
2006-03-27
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Structural and functional analyses of PL-7 alginate lyase2005
Author(s)
Kohei, Ogura, Masayuki, Yamasaki, Wataru, Hashimoto, Bunzo, Mikami, Kousaku, Murata
Organizer
Annual Meeting of The Society for Biotechnology
Place of Presentation
Tsukuba International Congress Center, Japan
Year and Date
2005-11-16
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Similarity in catalytic reaction mechanism of alginate lyases with different structural scaffold2005
Author(s)
Masayuki, Yamasaki, Kohei, Ogura, Wataru, Hashimoto, Kousaku, Murata, Bunzo, Mikami
Organizer
Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, Kansai Branch
Place of Presentation
Osaka University
Year and Date
2005-09-30
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Creation of superbacteria with high dioxin-degrading ability through transplantation of bacterial superchannel2005
Author(s)
Yuji, Aso, Yukiko, Miyamoto, Wataru, Hashimoto, Kousaku, Murata
Organizer
Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, Kansai Branch
Place of Presentation
Osaka University
Year and Date
2005-09-30
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Metabolism of alginate oligosaccharides in Agrobacterium tumefaciens C582005
Author(s)
Akihito, Ochiai, Osamu, Miyake, Wataru, Hashimoto, Kousaku, Muratra
Organizer
Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, Kansai Branch
Place of Presentation
Osaka University
Year and Date
2005-09-30
Description
「研究成果報告書概要(欧文)」より
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