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

一些致病性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。构成细胞表面基质一部分的哺乳动物糖胺聚糖是微生物酶的典型靶标。不饱和糖基水解酶（UGL），该水解酶在遗传上首次在杆菌中鉴定出来。菌株GL1催化通过基质降解多糖酶（例如透明质酸盐和软骨素溶酶酶）的反应产生的不饱和尿酸的水解释放，这表明这些酶在微生物感染中起着巨大的作用。 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 hydrolonate lyase homologue (xanthan lyase) were clarified. A single tyrosine … More retention abstracts C5-proton of the uronate retention 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 anime configuration, UGL uniquely triggers hydrolysis of vinyl ether groups in the unsaturated uronate retention but not of the糖苷键。最终的完整基因组序列分析表明，从细菌到真菌（超过70种）的大量微生物在其基因组中具有一个同源基因。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系列GH-88。 UGL的微生物生产者包括致病细菌，例如梭状芽胞杆菌，链球菌和振动。尤其是在链球菌，agalactiae链球菌，肺炎链球菌，肺炎链球菌和苏伊斯链球菌中发现的酶同源基因。与ugl芽孢杆菌不同，链球菌UGL在GalNAC的C4上与不饱和软骨蛋白二糖一起起作用。较少的

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

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

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

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

微生物による海洋資源アルギン酸のバイオエネルギーへの変換

微生物将海洋资源海藻酸转化为生物能源

• 发表时间: 2008
• 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
• 通讯作者: 村田幸作