Molecular basis for cell surface architecture and evolution of bacterial flagellin by structure and unction analysis of the protein

通过蛋白质的结构和功能分析，了解细胞表面结构和细菌鞭毛蛋白进化的分子基础

• 批准号: 17380053
• 负责人: MURATA Kousaku
• 金额: $ 9.93万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17380053/
• 关键词: Sphingomonas; Alginate; Flagellin; Flagellum; Receptor; X-ray crystallography; Pit; T4 phage; フィンガードメイン; 鞭毛タンパク質; 細菌細胞表層構造; Sphingomonas属細菌; 鞭毛進化; 細胞表層; 襞構造

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

海藻酸同化鞘单胞菌菌株Al通过细胞表面褶分子的重排形成一个口状的坑，并通过坑吸收大分子海藻酸盐，坑的功能是细胞外多糖的浓缩器。在海藻酸盐存在的情况下，两个蛋白p5 （40 kDa）和p6 （31 kDa）在细胞表面被诱导表达，这表明它们负责坑的形成。本研究分析了p5和p6的结构和功能。p5和p6都类似于细菌鞭毛蛋白，尽管菌株Al不形成鞭毛。一种p6干扰物在海藻酸盐培养基中显示出明显的生长迟缓，p5和p6的双重破坏导致生长失败，这表明这些鞭毛蛋白同源物可能对细胞生存至关重要。p6干扰物的细胞表面与野生菌株Al的不同之处在于，细胞表面凹坑的形成不完全，细胞表面结构由褶状结构转变为网状结构。表面等离子体共振生物传感器分析表明，这两种蛋白都具有高亲和力（解离常数Kd = ~ nM）特异性结合海藻酸盐。基于这些结果，定位于细胞表面的p5和p6可以被鉴定为外源海藻酸盐受体。p5由N-/ c端α (α1 + α2)结构域和中央B结构域组成。由于缺乏α_1结构域的p5突变体没有与海藻酸盐结合的能力，α_1结构域是与海藻酸盐结合所必需的。p5与沙门氏菌鞭毛蛋白比较，α (α1 + α2)结构域结构相似，说明细菌鞭毛蛋白中常见的海藻酸结合能力是由于一个结构域的保守性。另一方面，p5 β结构域在结构上与T4噬菌体蛋白gp11的手指结构域相似，位于基板与尾纤维之间的铰链区。手指结构域与基板蛋白gp10的结合对于尾部纤维在细菌细胞表面的附着很重要。手指结构域显示出与其他蛋白的亲和力，表明p5 β结构域作为锚点与菌株Al细胞表面分子相互作用，并有助于海藻酸结合结构域与外部环境的定向。少

项目成果

Creation of superbacteria for bioremediation through transplantation of bacterial superchannel

通过移植细菌超级通道创建用于生物修复的超级细菌

• 发表时间: 2005
• 作者: Yukiko;Miyamoto;Yuji;Aso;Wataru;Hashimoto;Kousaku;Murata
• 通讯作者: Murata

スーパー細菌の創成とバイオレメディエーションへの利用

超级细菌的创造及其用于生物修复

• 发表时间: 2006
• 期刊: 資源環境対策 42
• 作者: Wataru;Hashimoto;Yukie;Maruyama;Yuji;Aso;Kousaku;Murata;Zhongli Cui;Akihito Ochiai;Akihito Ochiai;Yukie Maruyama;Wataru Hashimoto;橋本渉;Chikako Fukuda;河井重幸;Akihito Ochiai;Karen Mine Harada;Yuji Aso;宮本裕希子;麻生祐司
• 通讯作者: 麻生祐司

A structural factor responsible for substrate recognition by Bacillus sp. GL1 xanthan lyase that acts specifically on pyruvated side chains of xanthan.

• DOI: 10.1021/bi0619775
• 发表时间: 2007-01
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Y. Maruyama;B. Mikami;W. Hashimoto;K. Murata

Superchannel of Bacteria: Biological Significance and New Horizons

• DOI: 10.1271/bbb.70635
• 发表时间: 2008-02
• 期刊: Bioscience, Biotechnology, and Biochemistry
• 作者: K. Murata;S. Kawai;B. Mikami;W. Hashimoto

Hydration of vinyl ether groups by unsaturated glycoside hydrolases and their role in bacterial pathogenesis.

• 发表时间: 2007-12
• 期刊: International microbiology : the official journal of the Spanish Society for Microbiology
• 作者: W. Hashimoto;T. Itoh;Y. Maruyama;B. Mikami;K. Murata