XAS STUDIES OF METAL TRANSFER

金属转移的 XAS 研究

• 批准号: 8170197
• 负责人: Ninian J Blackburn
• 金额: $ 0.34万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170197
• 关键词: Binding; Binding Sites; Chemistry; Computer Retrieval of Information on Scientific Projects Database; Copper; Data; Elements; Funding; Goals; Grant; Institution; Kinetics; Label; Ligand Binding; Location; Measures; Metals; Molecular Chaperones; Monitor; Process; Proteins; Reaction; Research; Research Personnel; Resources; Selenocysteine; Selenomethionine; Source; Spectrum Analysis; United States National Institutes of Health; Work; absorption; cytochrome c oxidase; interest; meetings; member; periplasm

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. Detecting and quantifying metal transfer reactions between chaperones and target proteins is currently of great interest. An important experimental requirement is the ability to label one member of the pair in order to track the location of the bound metal as the transfer proceeds. In this proposal we develop a strategy in which selenomethionine (SeM) or selenocysteine (Sec) are substituted for the native Met or Cys ligands of the binding sites of one member of the chaperone target pair. The Se label can then be used as a direct spectroscopic probe of the location of a copper atom during the transfer process via monitoring of the Se-Cu interaction by x-ray absorption spectroscopy at both the Se and Cu absorption edges. Our studies will include copper transfer reactions between (i) the periplasmic efflux proteins CusF and CusB, (ii) T thermophilus cytochrome oxidase CuA and Sco or PCuAC, and (iii) the mammalian CTR1 importer and hCCS. The goal of the work is to obtain evidence for selective transfer chemistry, measure the kinetics of the process, and use this data to interrogate the essential structural elements of the transfer mechanism.

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。 目前，检测和量化伴侣和靶蛋白之间的金属转移反应引起了极大的兴趣。一个重要的实验要求是能够标记一对成员的一个成员，以便在传输进行时跟踪绑定金属的位置。在此提案中，我们制定了一种策略，在该策略中，将硒甲米氨酸（SEM）或硒代半胱氨酸（SEC）取代代替伴侣靶靶对的一个成员的结合位点的天然MET或CYS配体。然后，可以通过在SE和CU吸收边缘通过X射线吸收光谱监测SE-CU相互作用，将SE标签用作传输过程中铜原子位置的直接光谱探针。我们的研究将包括（i）（i）（i）周围外排蛋白CUSF和CUSB，（ii）T嗜热细胞色素氧化酶CuA和SCO或PCOAC之间的铜转移反应，以及（iii）哺乳动物CTR1进口基和HCC。这项工作的目的是获得选择性转移化学的证据，测量过程的动力学，并使用此数据来询问转移机制的基本结构元素。