NMR Studies on the Structure and Dynamics of Glycan-Mediated IgG Interactions

聚糖介导的 IgG 相互作用的结构和动力学的 NMR 研究

• 批准号: 8232940
• 负责人: Adam Wesley Barb
• 金额: $ 4.07万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2012-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8232940
• 关键词: Affect; Age; Anti-Inflammatory Agents; Anti-inflammatory; Autoimmune Diseases; Binding; Biological; Carbohydrates; Cell Surface Receptors; Characteristics; Complex; Coupling; Crystallography; Dendritic Cells; Development; Disaccharides; Disease; Drug Design; Employee Strikes; Fc Immunoglobulins; Future; Galactose; Glycoproteins; Goals; Hand; Health; Human; Immunoglobulin G; Individual; Inflammatory; Joints; Knowledge; Label; Link; Location; Lymphocyte antigen CD50; Malignant Neoplasms; Measurement; Measures; Mediating; Methods; Modeling; Modification; Molecular; Molecular Conformation; Nuclear; Nuclear Magnetic Resonance; Play; Polysaccharides; Post-Translational Protein Processing; Process; Productivity; Protein Conformation; Protein Glycosylation; Proteins; Research; Residual state; Rheumatoid Arthritis; Role; Sampling; Sialic Acids; Structure; Techniques; Testing; Therapeutic; Tissues; X ray diffraction analysis; X-Ray Diffraction; base; conformer; design; effective therapy; extracellular; link protein; novel; protein protein interaction; public health relevance; receptor; receptor binding; sialylation; small molecule; tool

DESCRIPTION (provided by applicant): N-linked protein glycosylation is the most common eukaryotic protein modification and many diseases including autoimmune disorders and cancer are associated with compositional changes of N-glycans. A broad spectrum of N-glycans are specifically recognized by cell surface receptors, and these interactions likely play a critical role in these diseases. The long-term goals of this research are to explore the structure and dynamics of N-glycans as attached to a protein and as specifically coordinated in a protein-protein interaction. Rheumatoid arthritis (RA), an autoimmune disorder, is a crippling disease that can strike at any age, disabling and disfiguring its victims by destroying joint tissue. A recent link between RA and a specific posttranslationally modified form of immunoglobulin G (IgG) provides an important key to understanding the disease and finding a more effective treatment. The modification involves terminal sialylation of the biantennary N-glycans attached to the Fc fragment of IgG. This specific modification is found to dramatically increase the anti-inflammatory activity of IgG, suggesting molecular mimics of the sialylated Fc glycans may be a promising new treatment for RA. The aims of this proposal are: 1) To study the molecular character of terminal sialic acid residues and the effect of sialylation on the N-glycans associated with the Fc fragment of IgG using nuclear magnetic resonance (NMR)-based techniques; 2) To develop techniques for the assignment of N-linked glycan resonances. This will utilize the 13C-labeling strategy of Aim 1 combined with a novel mass spectral analysis technique to correlate resonances with either the alpha1-3 or alpha1-6 branch of these complex-type N-glycans; 3) To calculate a structure and evaluate the dynamics of the sialic acid-galactose disaccharides at the termini of the N-glycan; and 4) To measure the interaction of the sialylated Fc fragment with its proposed receptor, dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN). PUBLIC HEALTH RELEVANCE: Rheumatoid arthritis is a debilitating disease that limits the health and productivity of affected individuals. This proposal will utilize recent scientific discoveries to study the details of molecular interactions underlying rheumatoid arthritis. This knowledge will guide the future development of more effective treatments of rheumatoid arthritis.

描述(申请人提供)：N-连接蛋白糖基化是最常见的真核蛋白质修饰，许多疾病，包括自身免疫性疾病和癌症，与N-糖链的组成变化有关。广泛的N-糖链是由细胞表面受体特异性识别的，这些相互作用可能在这些疾病中发挥关键作用。这项研究的长期目标是探索N-葡聚糖在蛋白质上的结构和动力学，以及在蛋白质-蛋白质相互作用中的特定协调。类风湿性关节炎(RA)是一种自身免疫性疾病，是一种可以在任何年龄发作的致残性疾病，通过破坏关节组织使患者残疾和毁容。最近在RA和一种特定的翻译后修饰形式的免疫球蛋白G(IgG)之间的联系为了解这种疾病和找到更有效的治疗方法提供了重要的关键。修饰包括连接在免疫球蛋白Fc片段上的二天线N-糖链的末端唾液酸化。这种特异性的修饰显著提高了免疫球蛋白的抗炎活性，提示唾液酸化的Fc多糖的分子模拟物可能成为治疗类风湿关节炎的一种有前途的新方法。这项建议的目的是：1)利用基于核磁共振的技术研究末端唾液酸残基的分子特征和唾液酸化对与免疫球蛋白Fc片段相关的N-糖链的影响；2)发展N-连接的糖链共振峰的分配技术。这将利用AIM 1的13C标记策略结合新的质谱分析技术来关联这些复合型N-聚糖的α1-3或α1-6分支的共振；3)计算N-糖链末端的唾液酸-半乳糖二糖的结构并评估其动力学；以及4)测量唾液酸化的Fc片段与其建议的受体-树突状细胞特异性ICAM-3-抓取非整合素(DC-SIGN)的相互作用。 公共卫生相关性：类风湿性关节炎是一种使人衰弱的疾病，它限制了受影响个人的健康和生产力。这项建议将利用最近的科学发现来研究类风湿性关节炎背后的分子相互作用的细节。这一认识将指导未来开发更有效的类风湿性关节炎治疗方法。