H/D EXCHANGE STUDIES OF NIKR

NIKR 的 H/D 交换研究

• 批准号: 7953910
• 负责人: PETER T CHIVERS
• 金额: $ 0.12万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7953910
• 关键词: Affinity; Bacteria; Binding; Biochemical; Biological Assay; Biological Process; Cells; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Equilibrium; Escherichia coli; Fluorescence; Funding; Gene Expression; Genes; Goals; Grant; Growth; Helicobacter pylori; Institution; Ions; Ligand Binding; Mass Spectrum Analysis; Microbe; Molecular; Molecular Chaperones; Molecular Genetics; Nickel; Operon; Pathway interactions; Property; Proteins; Regulation; Reporter; Research; Research Personnel; Resources; Source; Spectrum Analysis; Trace metal; United States National Institutes of Health; biomedical resource; in vivo; interest; protein function; trafficking; uptake

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Chivers lab is interested in how cells control intracellular levels of essential trace metals. A balance must be achieved between levels that are sufficient for growth and those that are toxic to the cell. They focus on the regulation and intracellular trafficking of nickel in microbes, in particular Escherichia coli and Helicobacter pylori. In these bacteria, nickel-enzymes are synthesized with the help of specific nickel-binding chaperones. The nickel-dependent transcriptional regulator NikR binds free nickel ions and represses expression of nickel uptake genes. NikR has also been shown to postively regulate gene expression of at least one operon (urrease) in H. pylori. The function of NikR indicates that free nickel ions accumulate only after sufficient nickel has been acquired to synthesize nickel-dependent enzymes. The high-affinity of NikR for nickel ions, however, suggests that nickel-trafficking pathways must actively compete with NikR for newly acquired nickel ions. A goal is to study the mechanism by which this partitioning is achieved. Our approach combines biochemical and biophysical studies of purified proteins (for example, fluorescence and UV-visible spectroscopy) with in vivo studies of protein function (molecular genetics, reporter assays) to correlate molecular properties, such as ligand-binding affinity, with biological function. A new direction is to use mass spectrometry, especially some of the biophysical approaches developed in the Michael Gross lab.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 Chivers Lab对细胞如何控制基本痕量金属的细胞内水平感兴趣。必须在足以生长的水平和对细胞有毒的水平之间达到平衡。他们专注于微生物中镍的调节和细胞内运输，特别是大肠杆菌和幽门螺杆菌的调节。在这些细菌中，镍酶是在特定的镍结合伴侣的帮助下合成的。依赖镍的转录调节剂NIKR结合游离镍离子并抑制镍摄取基因的表达。 NIKR还显示出在幽门螺杆菌中至少一个操纵子（urrease）的基因表达。 NIKR的功能表明，仅在获得足够的镍以合成镍依赖性酶的情况下，游离镍离子才积累。然而，NIKR的高亲和力是镍离子的高亲和力表明，流浪货物的途径必须与Nikr积极竞争新获得的镍离子。一个目标是研究实现这种分区的机制。我们的方法结合了纯化蛋白（例如荧光和紫外可见光谱）的生化和生物物理研究与蛋白质功能的体内研究（分子遗传学，报告基因分析）相关的分子特性，例如配体结合亲和力，与生物学功能相关。 一个新的方向是使用质谱法，尤其是在迈克尔·格罗斯（Michael Gross）实验室中开发的一些生物物理方法。