NEW NMR METHODOLOGY FOR CHARACTERIZING CARBOHYDRATE-PROTEIN INTERACTIONS

表征碳水化合物-蛋白质相互作用的新核磁共振方法

• 批准号: 7721569
• 负责人: JAMES H. PRESTEGARD
• 金额: $ 14.04万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721569
• 关键词: Amides; Amino Acids; B-Cell Activation; Carbohydrates; Cell Aggregation; Cell Differentiation process; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Disease; Environment; Funding; Grant; Inflammation; Institution; Label; Ligand Binding; Ligands; Malignant Neoplasms; Methodology; Modeling; Nuclear Magnetic Resonance; Numbers; Object Attachment; Oligosaccharides; Process; Protein-Carbohydrate Interaction; Proteins; Range; Relaxation; Research; Research Personnel; Residual state; Resources; Retrieval; ST6Gal I; Sialyltransferases; Signal Transduction; Site; Source; Structural Models; Transferase; United States National Institutes of Health; Virus Diseases; Work; analog; drug development; nuclear Overhauser enhancement; research study; tool; vector

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The formation of carbohydrate-protein complexes is important in a variety of processes involving the interaction of a cell with its environment. These processes include natural ones such as cell differentiation, cell aggregation, and cell signaling; they also include disease processes such as viral infection, malignancy, and unwanted inflammation. The development of drugs to moderate natural processes and inhibit disease processes begins with accurate structural models for oligosaccharide-protein interactions. These models have been difficult to get by traditional Nuclear Magnetic Resonance (NMR) approaches because of the dearth of short-distance NOE contacts in oligosaccharide-protein complexes. We are developing a number of experiments and analysis tools that allow retrieval of structural information when traditional NMR approaches fail. This particular project combines a single amino acid labeling strategy developed in the Moremen group with two new types of NMR-observable structural data. One type comes from residual dipolar couplings of labeled 15N-1H pairs in various amide bonds. These give orientational constraints on bond vectors. The other type comes from long range paramagnetic spin relaxation perturbation of labeled sites by synthesized nitroxide-carrying analogs of protein ligands. These give long-range distance constraints between the bound ligand and the labeled sites. The sialyltransferase, ST6Gal I, has been the primary target motivating this work. This transferase was first cloned more than 17 years ago (Weinstein, Ujita, Paulson, Fed. Proceed., 44, 690-690, 1985), but it continues to resist attempts at structural characterization by traditional means. It is of importance in the activation of B-cells among other things, but it also is representative of many important mammalian proteins that are best expressed in non-bacterial hosts.

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