Molecular and structural biostudies on streptococcalinyasion/infection mechanisms to host cells through degradation of extracellular matrices glyoceaminoglycans

通过细胞外基质糖胺聚糖降解对宿主细胞链球菌感染/感染机制的分子和结构生物研究

• 批准号: 18580075
• 负责人: HASHIMOTO Wataru
• 金额: $ 2.48万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18580075/
• 关键词: Streptococcus; Glycosaminoalycan; Polysaccharide lyase; Unsaturated glucuronyl hydrolase; Invasion; infection mechanism; X-ray crystallography; Sulfated saccharide; Extracellular matrix; コンドロイチン; ヒアルロン酸リアーゼ; 遺伝子クラスター

Some pathogenic microorganisms produce polysaccharide-degrading enzymes (lyases and hydrolases) to invade mammalian and/or plant cells. Mammalian glycosaminoglycans that form part of cell surface matrix are typical targets for microbial enzymes. Unsaturated glucuronyl hydrolase (UGL), which was genetically first identified in Bacillus sp. strain GL1, catalyzes the hydrolytic release of an unsaturated uronic acid from oligosaccharides produced through the reaction of the matrix-degrading polysaccharide lyases (e.g., hyaluronate and chondroitin lyases), suggesting that these enzymes function as a virulent factor in microbial infection. In this study, structure and function relationship of glycosaminoglycan-degrading enzymes, lyase and hydrolase, was analyzed.Based on X-ray crystallography of enzyme-substrate complexes and site-directed mutagenesis, mechanisms for catalytic reaction and substrate recognition of hyaluronate lyase homologue (xanthan lyase) were clarified. A single tyrosine … More residue abstracts C5-proton of the uronate residue at subsite +1 and donates the proton to glycosidic bond to be cleaved.In contrast with general glycoside hydrolases with the retention or inversion catalytic mechanism of an anomeric configuration, UGL uniquely triggers hydrolysis of vinyl ether groups in the unsaturated uronate residue but not of the glycosidic bond.Recent complete genome sequence analyses indicate that a large number of microorganisms ranging from bacteria to fungi (over 70 species) have a UGL homologous gene in their genome. In the Carbohydrate-Active enZyme (CAZy) database, UGL and its homologues form a new family, GH-88. Microbial producers of UGL include pathogenic bacteria such as clostridia, streptococci, and vibrios. The enzyme homologous gene has been found especially in streptococci, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, and Streptococcus suis. Unlike the bacillus UGL, streptococcal UGL acts on unsaturated chondroitin disaccharide with a sulfate group at C4 of GalNAc. Less

一些病原微生物产生多糖降解酶（裂解酶和水解酶）以侵入哺乳动物和/或植物细胞。哺乳动物糖胺聚糖形成细胞表面基质的一部分，是微生物酶的典型靶标。不饱和葡糖醛酸水解酶（UGL）在遗传上首先在芽孢杆菌属菌株GL 1中鉴定，其催化不饱和糖醛酸从通过基质降解多糖裂解酶（例如，透明质酸和软骨素裂解酶），表明这些酶在微生物感染中起毒性因子的作用。本研究分析了糖胺聚糖降解酶（裂解酶和水解酶）的结构与功能关系，并基于酶-底物复合物的X射线晶体学和定点突变技术，阐明了透明质酸裂解酶同系物（黄原胶裂解酶）的催化反应和底物识别机制。单个酪氨酸 ...更多信息 残基在亚位点+1处夺取糖醛酸残基的C5-质子，并将该质子提供给待裂解的糖苷键。与具有异头构型的保留或转化催化机制的一般糖苷水解酶相比，UGL独特地引发不饱和糖醛酸残基中的乙烯基醚基团的水解，但不引发糖苷键的水解。从细菌到真菌（超过70个物种）在它们的基因组中具有UGL同源基因。在碳水化合物活性酶（CAZy）数据库中，UGL及其同源物形成了一个新的家族GH-88。UGL的微生物生产者包括病原菌，如梭菌、链球菌和弧菌。该酶的同源基因尤其在链球菌、无乳链球菌、肺炎链球菌、化脓链球菌和猪链球菌中发现。与芽孢杆菌UGL不同，链球菌UGL作用于在GalNAc的C4处具有硫酸酯基的不饱和软骨素二糖。少

项目成果

Structure of unsaturated rhamnogalacturonyl hydrolase complexed with substrate

与底物复合的不饱和鼠李糖半乳糖醛酸水解酶的结构

• 发表时间: 2006
• 期刊: Biochemical and Biophysical Research Communications 347
• 作者: Yukie;Maruyama;Bunzo;Mikami;Wataru;Hashimoto;Kousaku;Murata;Zhongli Cui;Akihito Ochiai;Akihito Ochiai;Yukie Maruyama;橋本 渉;Magdy Mahfouz;Anita Chaudhari;Yukie Maruyama;Takafumi Itoh
• 通讯作者: Takafumi Itoh

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

連鎖球菌による宿主細胞外マトリクス・グリコサミノグリカンの分解

链球菌对宿主细胞外基质糖胺聚糖的降解

• 发表时间: 2007
• 作者: Takafumi;Itoh;Bunzo;Mikami;Wataru;Hashimoto;Kousaku;Murata;落合秋人;橋本 渉;伊藤貴文;丸山如江;村田幸作;宮本裕希子;落合秋人;川眞田明子;丸山如江;落合秋人;橋本 渉
• 通讯作者: 橋本 渉

Crystal structure of unsaturated glucuronyl hydrolase complexed with substrate : Molecular insights into its catalytic reaction mechanism

与底物复合的不饱和葡萄糖醛酸水解酶的晶体结构：对其催化反应机制的分子见解

• 发表时间: 2006
• 期刊: Journal of Biological Chemistry 281
• 作者: Takafumi;Itoh;Wataru;Hashimoto;Bunzo;Mikami;Kousaku;Murata
• 通讯作者: Murata

サルモネラ菌由来機能不明タンパク質YihSの構造・機能解析:ポスト構造ゲノミクスに向けて

YihS（一种源自肠沙门氏菌的功能未知的蛋白质）的结构和功能分析：迈向后结构基因组学

• 发表时间: 2008
• 作者: Takafumi;Itoh;Wataru;Hashimoto;Bunzo;Mikami;Kousaku;Murata;Takafumi Itoh;Takafumi Itoh;Takafumi Itoh;Akihito Ochiai;Takafumi Itoh;伊藤貴文
• 通讯作者: 伊藤貴文