STRUCTURE OF MACROPHAGE CHEMOTACTIC ACTIVATING PROTEINS

巨噬细胞趋化激活蛋白的结构

• 批准号: 2073729
• 负责人: Tracy M Handel
• 金额: $ 11.83万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2073729
• 关键词: acidity /alkalinity; chemical stability; chemoattractants; chemotaxis; chimeric proteins; circular dichroism; cytokine; cytokine receptors; macrophage activating factor; molecular cloning; mutant; nuclear magnetic resonance spectroscopy; protein folding; protein structure function; receptor binding; site directed mutagenesis; stable isotope double label; structural biology; temperature; thermodynamics

The long term objective of the proposed research is to enhance our understanding of the molecular mechanisms of lymphocyte chemotaxis. This is a process critical to the normal functioning of the immune system, but also implicated in a variety of diseases such as atherosclerosis and autoimmune disease where chemotactic signaling becomes unbalanced. A growing family of small soluble chemokines has recently been discovered whose primary role is to direct the migration of specific lymphocytes to areas of physiological insult, and to activate the lymphocytes for destruction of the antigen. This occurs by the interaction of these chemokines with receptors on macrophages, neutrophils and T-cells which initiates a signal transduction cascade coordinating the inflammatory response. Understanding the detailed molecular structures of the chemokines, points of contact with the receptor binding sites, and regions of the proteins critical for signal transduction will advance our general understanding of lymphocyte chemotaxis. Detailed comparison of the structures of several related chemokines will also forward our understanding of protein folding and the energetic interactions which allow different primary sequences to adopt similar 3-dimensional structures. Accordingly, the specific aims of the project are to: 1) determine the dynamics and high resolution NMR structures of several homologous beta-chemokines (MCP-1, MIP-1alpha and I-3090 involved in macrophage-mediated chemotaxis and chronic inflammation. 2) evaluate the aggregation states and stabilities of MCP-1, MIP-1alpha and I-309 under a variety of solution conditions. 3) map the regions of the chemokines (MCP-1 and MIP-1alpha) important for receptor binding and signal transduction; this will be accomplished using mutagenesis and assays of functional biological activity (binding, chemotaxis, and Ca+2 influx).

拟议研究的长期目标是增强我们的 了解淋巴细胞趋化性的分子机制。 这 是对免疫系统正常功能至关重要的过程，但是 还涉及多种疾病，例如动脉粥样硬化和 自身免疫性疾病，趋化信号传导变得不平衡。一个 最近发现了不断增长的小型可溶性趋化因子 其主要作用是指导特定淋巴细胞的迁移到 生理侮辱区域，并激活淋巴细胞 抗原的破坏。 这是由于这些的相互作用而发生的 与受体趋化因子在巨噬细胞，中性粒细胞和T细胞上 启动信号转导级联协调炎症 回复。 了解详细的分子结构 趋化因子，与受体结合位点的接触点和区域 信号转导至关重要的蛋白质将推动我们的一般性 了解淋巴细胞趋化性。 详细比较 几种相关趋化因子的结构也将转发我们的 了解蛋白质折叠和允许的能量相互作用 不同的主要序列采用相似的3维结构。 因此，该项目的具体目的是： 1）确定几个的动力学和高分辨率NMR结构 同源β-化学因子（MCP-1，MIP-1Alpha和I-3090参与 巨噬细胞介导的趋化性和慢性炎症。 2）评估MCP-1，MIP-1Alpha的汇总状态和稳定性 和I-309在各种溶液条件下。 3）绘制趋化因子（MCP-1和MIP-1Alpha）的区域 受体结合和信号转导；这将使用 诱变和功能生物学活性的测定（结合， 趋化性，Ca+2涌入）。