THE STRUCTURE OF CALBINDIN-D28K

CALBINDIN-D28K 的结构

• 批准号: 6696610
• 负责人: RAJIV KUMAR
• 金额: $ 17.76万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-15 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6696610
• 关键词: calbindin; calcium ion; conformation; electrospray ionization mass spectrometry; fluorescence spectrometry; nuclear magnetic resonance spectroscopy; protein binding; protein folding; protein structure function; structural biology

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.

描述（改编自申请人的摘要）：Calbindin-D28K是一个 未知三级结构的生物学上必不可少的钙结合蛋白 这是正常神经功能所必需的。该蛋白质还发挥 在肠道中钙的运输中的重要作用 和肾脏。 该赠款应用的目的是确定解决方案第三级 Calbindin-D28K在无钙和钙结合形式的结构 高分辨率的核磁共振（NMR）光谱法确定 钙量增加时发生了什么结构变化 到蛋白质，从而获得对作用机理的见解 蛋白质。 该赠款应用的假设是Calbindin-D28K，一种生物学上 必需的，EF手，钙结合蛋白具有独特的三级结构 在结合钙时会发生构象变化。 具体目的是：（1）使用高分辨率的核磁 共振（NMR）光谱，溶液三级结构 Calbindin-D28K的无钙和钙饱和形式。 （2）至 确定钙结合后发生的顺序结构变化 到Calbindin-D28K。 （3）确定特定缺失的影响 Calbindin-D28K中的EF手在结构和钙结合特性上 Calbindin-D28K。 要使用的技术包括：高分辨率1H，15N和13C核 全长和突变钙蛋白的磁共振光谱； 电喷雾电离质谱；灯笼荧光 光谱；以及近乎和固有的蛋白质荧光测定 遥远的紫外线。 该项目具有重要意义，因为Calbindin-D28K的三级结构 在存在和不存在钙的情况下，初始的影响 钙对全球蛋白质结构的结合仍有待确定，并且 有关蛋白质折叠如何提供有关其作用的信息的信息 在细胞内。这些发现可能在 了解神经退行性疾病，小脑功能和长期 海马增强。

项目成果

Secreted frizzled-related protein 4 is a potent tumor-derived phosphaturic agent.

分泌型卷曲相关蛋白 4 是一种有效的肿瘤源性磷酸盐制剂。

• DOI: 10.1172/jci18563
• 发表时间: 2003
• 期刊: The Journal of clinical investigation
• 作者: Berndt,Theresa;Craig,TheodoreA;Bowe,AnnE;Vassiliadis,John;Reczek,David;Finnegan,Richard;JanDeBeur,SuzanneM;Schiavi,SusanC;Kumar,Rajiv
• 通讯作者: Kumar,Rajiv

Regulation of osteoblast growth by interactions between transforming growth factor-beta and 1alpha,25-dihydroxyvitamin D3.

通过转化生长因子-β 和 1α,25-二羟基维生素 D3 之间的相互作用调节成骨细胞生长。

• 发表时间: 2001
• 期刊: Critical reviews in eukaryotic gene expression
• 影响因子: 1.6
• 作者: Gurlek,A;Kumar,R
• 通讯作者: Kumar,R

The role of heat shock protein 70 in vitamin D receptor function.

热休克蛋白 70 在维生素 D 受体功能中的作用。

• DOI: 10.1006/bbrc.2001.4711
• 发表时间: 2001
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Lutz,W;Kohno,K;Kumar,R
• 通讯作者: Kumar,R

Distinctive dendritic cell modulation by vitamin D(3) and glucocorticoid pathways.

• DOI: 10.1016/s0006-291x(02)02262-3
• 发表时间: 2002-09
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: N. Xing;Monica L L Maldonado-Monica-L-L-Maldonado-15074187;L. Bachman;D. McKean;R. Kumar;M. Griffin

Emerging role of mass spectrometry in structural and functional proteomics.

质谱在结构和功能蛋白质组学中的新兴作用。

• DOI: 10.1016/s0065-3233(03)01021-0
• 发表时间: 2003
• 期刊: Advances in protein chemistry
• 作者: Naylor,Stephen;Kumar,Rajiv
• 通讯作者: Kumar,Rajiv