STRUCTURAL BIOLOGY OF EF-HAND CALCIUM BINDING PROTEINS

描述：钙和钙结合蛋白(CABPS)起着中枢作用 在细胞信号转导通路中的作用，并与 对健康和疾病有广泛的影响。在职能层面上， 细胞事件包括细胞生长和分化、细胞 细胞周期进程、细胞凋亡和代谢控制。这项研究计划 寻求对CaBP如何在分子水平上发挥作用的理解，以开发 能够控制绑定属性，并最终设计特定的 与之相关的生物活性和治疗策略 钙介导性疾病。 为了实现这一目标，三维结构和内部动力学 在存在和不存在钙离子的情况下测定特定的钙结合蛋白 使用核磁共振波谱。然后比较对结合的钙离子的反应 不同的是，它们的生物活性范围都有CaBP。一 我们的研究阶段涉及将CABP数据库的范围扩大到 钙调素。这种蛋白质是微管的重要组成部分。 在中心体的组织中心，这是准确所需的 细胞周期M(有丝分裂)阶段的染色体分离。 这是独一无二的特性。包括体内的磷酸化，显然是在 细胞周期依赖的方式，使其成为极具吸引力的目标 治疗性干预 但不同蛋白质之间的比较不足以解释为什么 以及为什么会出现功能和结构上的差异。这个级别的 理解需要对驱动力和 钙结合的分子细节及其引起的相关变化 蛋白质的结构和动力学。为了解决这些问题，第二阶段 我们的研究涉及Calbindin D9k，一种EF手CABP，已经被 以前所未有的细节水平为特点。该蛋白质的研究进展 结构和动力学，以尽可能高的分辨率组合 通过定点突变和蛋白质工程实验， 被用来拼凑出分子基础的完整图景 CA2亲和力、选择性、协作性及其传递 将特性结合到一系列不同的生物活性中。

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.