STRUCTURAL BIOLOGY OF EF-HAND CALCIUM BINDING PROTEINS

EF-Hand 钙结合蛋白的结构生物学

• 批准号: 6329694
• 负责人: WALTER J. CHAZIN
• 金额: $ 19.87万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6329694
• 关键词: bioenergetics; calbindin; calcium; calcium binding protein; chemical binding; computer graphics /printing; computer simulation; conformation; intermolecular interaction; molecular dynamics; mutant; nuclear magnetic resonance spectroscopy; physical model; protein structure function; site directed mutagenesis; structural biology

DESCRIPTION: Calcium and calcium-binding proteins, (CABPS) play a central role in cellular signal transduction pathways and are associated with a wide-range of effects on health and disease. At the functional level, Cal-mediated, cellular events include cell growth and differentiation, cell cycle progression, apoptosis and metabolic control. This research program seeks an understanding of how CaBPs work at the molecular level, to develop the ability to control binding properties and utlimately, to design specific biological activities and therapeutic strategies relevant to calcium-mediated disease. To attain this goaL the three-dimensional structures and internal dynamics of specific CaBPs are being determined in the presence and absence of Ca2+ using NMR spectroscopy. The responses to binding Ca2+ are then compared for different.CaBPs across the spectrum of their biological activities. One phase of our research involves extending the range of the CaBP database to caltracin. This protein is an essential component of the microtubule organizing center in the centrosome which is required for accurate chromosomal segregation during the M (mitosis) stage of the cell cycle. It's unique properties. including phosphorylation in-vivo apparently in a cell cycle-dependent manner, make it an extremely attractive target for therapeutic intervention But comparison between different proteins is not suffficient to explain how and why functional and structural differences occur. This level of understanding requires fundamental knowledge of the driving forces and molecular details of Ca2+ binding and the concomitant changes induced in protein structure and dynamics. To address these issues, the second phase of our research involves calbindin D 9k, an EF-hand CaBP that has been characterized at an unprecedented level of detail. Studies of the protein's structure and dynamics at the highest possible resolution, in combination with site-directed mutagenesis and protein engineering experiments, are being used to piece together a complete picture of the molecular basis for Ca2+ affinity, selectivity, cooperativity and the transmission of these binding properties into a diverse range of biological activities.

描述：钙和钙结合蛋白（CABPS）在体内起着重要的作用。 在细胞信号转导途径中的作用，并与 对健康和疾病的广泛影响。 在职能一级， Cal介导的细胞事件包括细胞生长和分化、细胞增殖和分化。 周期进展、凋亡和代谢控制。 这个研究项目 寻求了解CaBPs如何在分子水平上发挥作用， 能够控制绑定属性和可用性， 生物活性和治疗策略 钙介导的疾病 为了达到这一目标，三维结构和内部动力学 在存在和不存在Ca2+的情况下， 使用核磁共振光谱法。 然后比较对结合Ca2+的反应， 不同的。CaBP在其生物活性的范围内。 一 我们的研究阶段涉及扩展CaBP数据库的范围， caltracin。 这种蛋白质是微管的重要组成部分 中心体中的组织中心，这是准确 在细胞周期的M（有丝分裂）阶段染色体分离。 它的独特属性。 包括体内磷酸化， 细胞周期依赖的方式，使其成为一个非常有吸引力的目标， 治疗干预 但是不同蛋白质之间的比较不足以解释 以及为什么会出现功能和结构差异。 这种程度的 理解需要对驱动力的基本知识， Ca2+结合的分子细节和诱导的伴随变化， 蛋白质结构和动力学。 为了解决这些问题，第二阶段 我们的研究涉及钙结合蛋白D 9k，一种EF-手型CaBP， 以前所未有的详细程度来描述。 蛋白质的研究 结构和动力学在尽可能高的分辨率， 通过定点突变和蛋白质工程实验， 被用来拼凑一个完整的分子基础图片， Ca2+亲和力、选择性、协同性以及这些离子的传递 结合特性到各种生物活性中。