MINING THE ORAL MICROBIOME FOR NOVEL GLYCAN-BINDING MOLECULES

挖掘口腔微生物组中的新型聚糖结合分子

• 批准号: 8985451
• 负责人: Stefan Hans-Klaus Ruhl
• 金额: $ 33.37万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-05 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8985451
• 关键词: Adhesions; Affinity; Animals; Bacteria; Bacterial Adhesins; Bacterial Adhesion; Bacterial Polysaccharides; Binding; Binding Proteins; Bioinformatics; Biological; Carbohydrates; Cell surface; Communities; Dental; Detection; Environment; Exhibits; Foundations; Funding; Genomic approach; Genomics; Glycoproteins; Goals; Gram-Positive Bacteria; Grant; Habitats; Health; Human; Lectin; Ligand Binding; Mammals; Mediating; Medical; Microbial Biofilms; Mining; Mouth Diseases; Mucous body substance; N-glycolylneuraminic acid; Nature; Oral; Oral cavity; Oral health; Play; Polysaccharides; Property; Proteins; Recombinant DNA; Recombinants; Role; Salivary; Sampling; Sampling Studies; Serine; Specificity; Structure; Surface; Systemic disease; Techniques; Work; collaboratory; forging; member; novel; oral bacteria; oral biofilm; oral microbiome; receptor; research study; tool; tooth surface

DESCRIPTION (provided by applicant): In the oral environment, interactions between lectin-like adhesions on oral bacteria and complementary glycan recognition motifs on oral host cell and tooth surfaces and on the surfaces of other oral bacteria play important roles for initial bacterial colonization and the subsequent formation of multi-species biofilms that have the potential to turn into pathogenic habitats causing dental, oral, and perhaps systemic disease. Considering (a) the multitude of bacterial species identified in oral biofilms, (b) the rich diversty of glycans found on salivary glycoproteins, and (c) the innumerable glycan motifs expressed on the surface of other bacteria, it is likely that many more unidentified lectin-like bacterial adhesns exist in the oral microbiome, beyond the few ones currently known. Once identified, those bacterial lectin-like adhesins, many of which presumably will exhibit novel glycan-binding specificities, can be forged by recombinant DNA techniques into highly specific tools for the detecting and localization of corresponding glycan receptor structures. The overall objective of this application is to mine the oral microbiomes of humans and other mammals for novel glycan- binding bacterial adhesions by using glycan-functionalized targeted probes. Here, we focus on one well-characterized class of bacterial adhesions, the serine-rich repeat (SRR) proteins of Gram- positive bacteria. By identifying a novel set of glycan-binding proteins with unique ligand-binding properties, we aim to demonstrate as a proof of principle that it will become feasible to expand the range of currently available glycan-binding probes enormously. The specific aims of the project are to 1. Isolate SRR adhesion-bearing bacteria from the oral cavity of humans and other mammalian species that bind specifically to host sialoglycans containing 4- or 9-O-acetylated forms of N-acetyl and N-glycolylneuraminic acid. 2. Isolate SRR adhesin-bearing bacteria from human oral biofilms that bind specifically to bacterial polysaccharide receptors containing the unusual lectin-recognition motifs (Rhaα1-2Rha and Galα1- 6Glc). 3. Use bioinformatics to identify novel glycan-binding regions in SRR proteins, and use bacterial genomics to generate recombinant glycan-binding probes for detection of a diverse set of glycans.

描述(申请人提供)：在口腔环境中，口腔细菌上的凝集素样黏附与口腔宿主细胞和牙齿表面以及其他口腔细菌表面上的互补糖链识别基序之间的相互作用，对于初始细菌定植和随后形成多物种生物膜具有重要作用，这些生物膜有可能转变为致病生境，导致牙科、口腔甚至全身疾病。考虑到(A)在口腔生物膜中发现的大量细菌物种，(B)唾液糖蛋白上发现的丰富的葡聚糖多样性，以及(C)其他细菌表面表达的无数的葡聚糖基序，很可能在口腔微生物群中存在更多的未鉴定的凝集素样细菌粘附物，而不是目前已知的少数几个。一旦鉴定出来，这些细菌凝集素样粘附素，其中许多可能会表现出新的糖链结合特异性，可以通过重组DNA技术锻造成高度特异的工具，用于检测和定位相应的糖链受体结构。这项应用的总体目标是通过使用葡聚糖功能化的靶向探针来挖掘人类和其他哺乳动物的口腔微生物群，以寻找与糖结合的新型细菌粘连。在这里，我们专注于一类特征良好的细菌粘连，革兰氏阳性细菌的富含丝氨酸重复序列(SRR)蛋白。通过鉴定一组具有独特配体结合特性的新的糖链结合蛋白，我们的目的是证明作为原理的证据，极大地扩展目前可用的糖链结合探针的范围是可行的。该项目的具体目标是：1.从人类和其他哺乳动物的口腔中分离出与含有4-或9-O-乙酰化形式的N-乙酰和N-羟基神经氨酸的宿主唾液酸特异结合的SRR粘附菌。2.从含有特殊凝集素识别基序(Rhaα1-2Rha和Galα1-6Glc)的细菌多糖受体上分离出携带SRR粘附素的细菌。3.利用生物信息学鉴定SRR蛋白中新的糖链结合区，并利用细菌基因组学产生重组糖链结合探针，用于检测一系列不同的糖链。