THE STRUCTURE OF CALBINDIN-D28K

描述(改编自申请人摘要)：Calbindin-D28K是一种 未知三级结构的生物必需钙结合蛋白 这是正常神经功能所必需的。这种蛋白质还扮演着一个 在肠上皮细胞钙转运中的重要作用 还有肾脏。 这项赠款申请的目标是确定第三方解决方案 钙结合蛋白-D28K在无钙和钙结合状态下的结构 高分辨率核磁共振(核磁共振)谱，以确定 随着钙含量的增加，结合会发生什么样的结构变化 从而对蛋白质的作用机制有深入的了解 蛋白质。 这项拨款申请的假设是Calbindin-D28K，一种生物学上的 必需的EF-Hand钙结合蛋白，具有独特的三级结构 它与钙结合后发生构象变化。 具体目标是：(1)获得、利用高分辨率核磁 核磁共振波谱，溶液的三级结构 无钙和钙饱和形式的钙结合蛋白-D28K。(2)至 确定钙结合时发生的顺序结构变化 至Calbindin-D28K。(3)确定删除特定名称的影响 EF-Hands in calbindin-D28K对钙结合蛋白结构和钙结合性能的影响 钙结合蛋白-D28K。 将使用的技术包括：高分辨率1H、15N和13C核 全长和突变型钙结合蛋白的磁共振波谱； 电喷雾电离质谱仪；镧系元素荧光 光谱学；以及近端和近端的内源蛋白质荧光测定 远紫外线范围。 该项目具有重要意义，因为Calbindin-D28k的三级结构 在有无钙的情况下，最初的影响是未知的 全球蛋白质结构上的钙结合仍有待确定，以及 关于蛋白质如何折叠的信息可以提供对它如何作用的洞察 在细胞内。这些发现可能会在未来的 对神经退行性疾病、小脑功能和远期的认识 海马区增强。

DESCRIPTION (Adapted from the Applicant's Abstract): Calbindin-D28K is a biologically essential calcium-binding protein of unknown tertiary structure that is required for normal neural function. The protein also plays an important role in the transport of calcium in epithelial cells of the intestine and kidney. The objective of this grant application is to determine the solution tertiary structure of calbindin-D28K in calcium-free and calcium-bound forms by use of high resolution nuclear magnetic resonance (NMR) spectroscopy, to determine what structural changes occur upon the binding of increasing amounts of calcium to the protein, and to thereby gain insights into the mechanism of action of the protein. The hypothesis of this grant application is that calbindin-D28K, a biologically essential, EF-hand, calcium-binding protein, has a unique tertiary structure which undergoes conformational changes upon binding calcium. The specific aims are: (1) To obtain, using high-resolution nuclear magnetic resonance (NMR) spectroscopy, the solution tertiary structure of the calcium-free and the calcium-saturated forms of calbindin-D28K. (2) To determine the sequential structural changes which occur upon binding of calcium to calbindin-D28K. (3) To determine the effects of deletions of specific EF-hands in calbindin-D28K on the structure and calcium-binding properties of calbindin-D28K. The techniques to be used include: high resolution 1H, 15N and 13C nuclear magnetic resonance spectroscopy of the full-length and mutant calbindins; electrospray ionization mass spectrometry; lanthanide fluorescence spectroscopy; and intrinsic protein fluorescence determinations in the near and far UV-range. The project is of significance because the tertiary structure of calbindin-D28k in the presence and absence of calcium is unknown, the effects of initial calcium binding on global protein structure remain to be determined, and information on how the protein folds could provide insight into how it acts within cells. These findings will potentially be of significance in the understanding of neurodegenerative disorders, cerebellar function and long-term hippocampal potentiation.