Structural Interactions of Bacterial Adhesin with Glycosaminoglycans

细菌粘附素与糖胺聚糖的结构相互作用

基本信息

批准号：
8400895
负责人：
Xu Wang
金额：
$ 22.66万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8400895
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 Health Homology Modeling Human Immune system Infection Inorganic Sulfates Instruction Interdisciplinary Study Knowledge Learning Lipids Lipoproteins Lyme Disease Mammalian Cell Membrane Mentors Methodology Methods Microbe Modeling Modification Mutation Nature Phase Polysaccharides 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

项目摘要

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.

抗生素抗性正成为迫在眉睫的健康威胁。打击这一目标的一种策略是抑制微生物对人类细胞的粘附。但是，微生物与目标之间的特定结构相互作用细胞的表征没有很好，从而阻碍了新的抗微生物剂的发展。这提案努力通过对相互作用进行深入的结构研究来克服这种缺陷在细菌粘附素和糖胺聚糖（GAGS）之间，这是一种无处不在的多糖细胞表面和细菌粘附素的主要靶标。该提案的重点将放在装饰上结合蛋白（DBP），一种来自Borlelia burgdorferi的粘附蛋白，莱姆病的病因学药。学习 GAGS与DBP之间的相互作用，DBP的结构在细胞质及其天然脂蛋白中的结构形式将使用溶液NMR求解。将这些结构作为基础，DBP与将确定异质和均匀的插科打寡聚物。这项研究的主要目标是获取dbp的高分辨率结构，这些结构复合起来，从这些结构中推导可以针对靶向的GAG和DBP之间的相互作用，以防止细菌对人类细胞的粘附。最后，由于通常以蛋白聚糖的形式存在插科打g，因此将尝试表征 DBP与完整decorin之间的相互作用，DBP主要针对的蛋白聚糖。主要目标该提案的这一部分将是使用完整的Decorin构建DBP的复杂结构，仅利用 NMR得出的定向约束和完整decorin的二聚体性质。拟议的工作将在复杂的碳水化合物研究中心进行，这是一个多学科的研究中心研究所和该国主要的血糖研究中心之一。候选人的长期职业目标是了解插科打术与蛋白质目标的相互作用的规则并设计将这些相互作用转向所需方向的策略。候选人在指导阶段将是获得用于净化的化学和生化方法的熟练程度 GAG低聚物并学习使用哺乳动物表达系统，这对于获得完整至关重要 Decorin。这些技术的知识将使候选人能够在提议的区域。