Structural Interactions of Bacterial Adhesin with Glycosaminoglycans

细菌粘附素与糖胺聚糖的结构相互作用

• 批准号: 7713686
• 负责人: Xu Wang
• 金额: $ 7.82万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7713686
• 关键词: Address; Adhesions; Affinity; Age; Anti-Adhesion Agent; Antibiotic Resistance; Area; Bacteria; Bacterial Adhesins; Bacterial Adhesion; Binding; Binding Proteins; Binding Sites; Biochemical; Biological; Borrelia burgdorferi; Carbohydrates; Cell physiology; Cell surface; Cells; Chemicals; Chondroitin Sulfates; Complex; Core Protein; Country; Cytoplasmic Structures; Cytoplasmic Tail; Dermatan Sulfate; Development; Docking; Drug Formulations; Event; Foundations; Glycobiology; Glycopeptides; Glycoproteins; Glycosaminoglycans; Goals; Hand; Health; Homology Modeling; Human; Immune system; Infection; Inorganic Sulfates; Interdisciplinary Study; Knowledge; Learning; Lipids; Lipoproteins; Lyme Disease; Mammalian Cell; Membrane; Mentors; Methodology; Methods; Microbe; Modeling; Modification; Mutation; Nature; Phase; Polysaccharides; Principal Investigator; Process; Protein Binding; Protein Isoforms; Proteins; Proteoglycan; Public Health; Relaxation; Research; Research Institute; Resolution; Role; Signal Transduction; Skin; Solutions; Spin Labels; Structure; Supervision; Surface; System; Techniques; Tissues; Training; Universities; Unspecified or Sulfate Ion Sulfates; Vector-transmitted infectious disease; Work; antimicrobial; antimicrobial drug; base; career; cell motility; combat; decorin; decorin binding protein B; dermatan sulfate chondroitin sulfate; design; falls; pathogenic bacteria; prevent; protein complex; protein protein interaction; protein structure; restraint

DESCRIPTION (provided by applicant): Antibiotic resistance is becoming an imminent health threat. One strategy for combating this is to inhibit the adhesion of microbes to human cells. However, specific structural interactions between microbes and target cells are not well characterized, thus hampering the development of new anti-microbial agents. This proposal strives to overcome this deficiency by conducting an in-depth structural study of the interactions between bacterial adhesins and glycosaminoglycans (GAGs), a ubiquitous polysaccharide found on the surface of cells and a primary target for bacterial adhesins. The focus of the proposal will be on the decorin binding protein (dbp), an adhesin from Borrelia burgdorferi, the etiological agent of Lyme disease. To study the interactions between GAGs and dbp, the structures of dbp in both cytoplasmic and its native lipoprotein form will be solved using solution NMR. Using these structures as bases, dbp's interactions with heterogeneous and homogeneous GAG oligomers will be determined. The main goal of this research is to obtain high resolution structures of dbp complexed to GAG and deduce from these structures the specific interactions between GAG and dbp that can be targeted to prevent the adhesion of bacteria to human cells. Finally, as GAGs often exist in the form of proteoglycans, an attempt will be made to characterize the interactions between dbp and intact decorin, the proteoglycan that dbp primarily targets. The main goal for this portion of the proposal will be to construct the complex structure of dbp with intact decorin utilizing only NMR derived orientational restraints and the dimeric nature of intact decorin. The proposed work will be carried out at the Complex Carbohydrate Research Center, a multidisciplinary research institute and one of the country's premier glycobiology research centers. The candidate's long term career goal is to understand the rules governing the interactions of GAGs with its protein targets and devise strategies for steering these interactions in a desired direction. The candidate's immediate goal during the mentored phase will be to acquire proficiency with the chemical and biochemical methods used to purify GAG oligomers and learn the use of mammalian expression systems that will be crucial for obtaining intact decorin. The knowledge of these techniques will enable the candidate to conduct independent research in the proposed area. Relevance: The results of this proposal will be invaluable for the design of a new class of anti-microbial agents that specifically inhibits the adhesion of microbials to human cells. It also adds to our understanding of how sulfated polysaccharides interact with proteins to initiate important biological events.

描述（由申请人提供）：抗生素耐药性正在成为迫在眉睫的健康威胁。对抗这种情况的一种策略是抑制微生物对人体细胞的粘附。然而，微生物和靶细胞之间的特定结构相互作用没有得到很好的表征，从而阻碍了新的抗微生物剂的开发。该提案致力于通过对细菌粘附素和糖胺聚糖（GAG）之间的相互作用进行深入的结构研究来克服这一缺陷，糖胺聚糖是一种在细胞表面发现的普遍存在的多糖，也是细菌粘附素的主要靶标。该提案的重点将是核心蛋白聚糖结合蛋白（DBP），一种来自莱姆病病原体伯氏疏螺旋体的粘附素。为了研究GAG和DBP之间的相互作用，将使用溶液NMR解析胞质及其天然脂蛋白形式的DBP的结构。使用这些结构作为基础，将确定DBP与异质和同质GAG寡聚体的相互作用。本研究的主要目标是获得与GAG复合的DBP的高分辨率结构，并从这些结构中推断出GAG和DBP之间的特异性相互作用，这些相互作用可以靶向防止细菌粘附到人体细胞。最后，由于GAG通常以蛋白聚糖的形式存在，因此将尝试表征DBP与完整核心蛋白聚糖（DBP主要靶向的蛋白聚糖）之间的相互作用。这部分提案的主要目标是仅利用NMR衍生的取向限制和完整核心蛋白聚糖的二聚体性质构建具有完整核心蛋白聚糖的DBP的复杂结构。拟议的工作将在复杂碳水化合物研究中心进行，这是一个多学科研究机构，也是该国首屈一指的糖生物学研究中心之一。候选人的长期职业目标是了解GAGs与其蛋白质靶点相互作用的规则，并设计出将这些相互作用导向所需方向的策略。候选人在指导阶段的直接目标是熟练掌握用于纯化GAG寡聚体的化学和生物化学方法，并学习使用哺乳动物表达系统，这对于获得完整的核心蛋白聚糖至关重要。这些技术的知识将使候选人能够在拟议的领域进行独立的研究。 相关性：该提案的结果对于设计一类新的抗微生物剂，特别是抑制微生物对人体细胞的粘附是非常宝贵的。它还增加了我们对硫酸化多糖如何与蛋白质相互作用以引发重要生物事件的理解。