THE STRUCTURE OF CALBINDIN-D28K

描述（改编自申请人的摘要）：钙结合蛋白-D28 K是一种钙结合蛋白。 三级结构未知的生物必需钙结合蛋白 这是正常神经功能所必需的。这种蛋白质也起到了 在肠上皮细胞钙转运中的重要作用 和肾脏。 这项拨款申请的目的是确定解决方案， 无钙和钙结合形式钙结合蛋白-D28 K的结构 高分辨率核磁共振（NMR）光谱，以确定 当钙的结合量增加时， 蛋白质，从而获得洞察的作用机制， 蛋白质。 这项拨款申请的假设是，钙结合蛋白-D28 K，一种生物学上的 必需的EF-手，钙结合蛋白，具有独特的三级结构 其在结合钙时经历构象变化。 具体目标是：（1）利用高分辨核磁共振技术， 共振（NMR）光谱，溶液的三级结构 钙结合蛋白-D28 K的无钙形式和钙饱和形式。(2)到 确定钙结合后发生的连续结构变化 至钙结合蛋白-D28 K。(3)为了确定特定基因缺失的影响， EF-手在calbindin-D28 K上的结构和钙结合性质 钙结合蛋白-D28 K。 将使用的技术包括：高分辨率1H、15 N和13 C核 全长和突变钙结合蛋白的磁共振波谱; 电喷雾电离质谱;镧系荧光 光谱学;和内在蛋白质荧光测定在近和 远紫外线范围 该项目具有重要意义，因为Calbindin-D28 k的三级结构 在存在和不存在钙的情况下是未知的， 钙结合在整体蛋白质结构上仍有待确定， 关于蛋白质如何折叠的信息可以深入了解它是如何起作用的 在细胞内。这些发现将可能对 了解神经退行性疾病，小脑功能和长期 海马增强

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.