CHARACTERIZATION OF THE STRUCTURE OF A LEAD-DEPENDENT DNAZYME

铅依赖性脱氧核糖核酸酶的结构表征

• 批准号: 7722004
• 负责人: GERARD C WONG
• 金额: $ 0.31万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722004
• 关键词: 3-Dimensional; Adopted; Binding; Binding Sites; Buffers; Catalytic DNA; Catalytic Domain; Chemicals; Complex; Computer Retrieval of Information on Scientific Projects Database; Condition; Dependence; Environment; Fluorescence Resonance Energy Transfer; Funding; Grant; Institution; Ions; Knowledge; Metals; Process; Research; Research Personnel; Resources; Sodium Chloride; Source; Structure; System; United States National Institutes of Health; cofactor; lead concentration; lead ion; three dimensional structure

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. We have previously characterized the structure of the 17E DNAzyme using Fluorescence Resonance Energy Transfer (FRET). This study revealed that in the presence of Zn2+ the DNAzyme undergoes a two-step folding process in order to adopt a more compact structure. We propose to use EXAFS to further study the three-dimensional structure of the 17E catalytic core when bound to Pb2+. Pb has core-shell levels that are accessible using EXAFS. The catalytic activity of the Pb DNAzyme and the dependence of activity on Pb concentration and buffer conditions have been thoroughly characterized. The chemical environment around the Pb binding site is expected to be complex. However, spatial correlations in complex, noncrystalline systems are routinely studied using EXAFS. We intend to investigate the EXAFS of Pb in order to determine the distances between the Pb metal ion cofactor and key atoms in the DNAzyme catalytic core. The DNAzyme structure will be investigated as a function of Pb concentration, monovalent salt concentration, and pH. This 3-D information will then be combined with existing knowledge of the sequence and structure of the DNAzyme in order to form a better understanding of the functional structure of the DNAzyme.

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表示。列出的机构是 对于中心，这不一定是调查员的机构。 我们先前使用荧光共振能量转移（FRET）表征了17E Dnazyme的结构。 这项研究表明，在存在Zn2+的情况下，Dnazyme经历了两步折叠过程，以采用更紧凑的结构。 我们建议使用EXAFS进一步研究与PB2+结合的17E催化核心的三维结构。 PB具有使用EXAFS访问的核心壳级别。 Pb dnazyme的催化活性以及活性对PB浓度和缓冲条件的依赖性已彻底表征。 PB结合位点周围的化学环境预计将很复杂。 但是，使用EXAFS常规研究了复杂的非晶体系统中的空间相关性。 我们打算研究PB的EXAF，以确定DNAZYME催化核心中Pb金属离子辅因子和钥匙原子之间的距离。 Dnazyme结构将作为PB浓度，单价盐浓度和pH的函数研究。 然后，此3-D信息将与现有的Dnazyme序列和结构的知识相结合，以更好地理解对Dnazyme的功能结构。