NMR/MS METHODOLOGY FOR RESONANCE ASSIGNMENT OF CARBOHYDRATE PROCESSING PROTEINS

碳水化合物加工蛋白共振归属的 NMR/MS 方法

• 批准号: 7601108
• 负责人: JAMES H. PRESTEGARD
• 金额: $ 14.11万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601108
• 关键词: Amides; Carbohydrates; Cell Aggregation; Cell Differentiation process; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Disease; Environment; Funding; Galectin 3; Glycoproteins; Grant; Inflammation; Institution; Link; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Methodology; Methods; Modeling; Oligosaccharides; Process; Proteins; Protons; Rate; Research; Research Personnel; Resources; ST6Gal I; Sialyltransferases; Signal Transduction; Site; Source; Structural Models; United States National Institutes of Health; Virus Diseases; Work; drug development; novel strategies

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 the protein and for the oligosaccharide-protein interactions. NMR methods provide a potential source of these models, but a prerequisite is assignment of resonances to specific sites both within the protein and within the oligosaccharide. This has been particularly difficult for the large, difficult to express, proteins involved in carbohydrate processing. A good example of such a protein is the sialyltransferase, ST6Gal I, described in subproject 0007. We are developing a novel approach that links NMR resonances with mass spectrometry (MS) fragment identification through amide proton exchange rates measured by both methods. This work is beginning with applications to a simple model protein, Galectin-3, but will eventually target ST6Gal I and other glycoproteins.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 碳水化合物-蛋白质复合物的形成在涉及细胞与其环境相互作用的各种过程中非常重要。这些过程包括自然过程，例如细胞分化、细胞聚集和细胞信号传导；它们还包括病毒感染、恶性肿瘤和不必要的炎症等疾病过程。缓和自然过程和抑制疾病过程的药物的开发始于蛋白质和寡糖-蛋白质相互作用的精确结构模型。 NMR 方法提供了这些模型的潜在来源，但先决条件是将共振分配到蛋白质内和寡糖内的特定位点。对于参与碳水化合物加工的大的、难以表达的蛋白质来说，这尤其困难。这种蛋白质的一个很好的例子是唾液酸转移酶 ST6Gal I，在子项目 0007 中描述。我们正在开发一种新方法，通过两种方法测量的酰胺质子交换率将 NMR 共振与质谱 (MS) 片段鉴定联系起来。这项工作首先应用于简单的模型蛋白 Galectin-3，但最终将针对 ST6Gal I 和其他糖蛋白。