Steady-State and Stopped-flow Spectropolarimeter at NCSU

NCSU 的稳态和停流旋光仪

• 批准号: 7038005
• 负责人: ALLAN CLAY CLARK
• 金额: $ 22.19万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2007-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7038005
• 关键词: RNA; circular dichroism; nucleic acids; proteins

DESCRIPTION (provided by applicant): Three NCSU scientists working in fields in which protein-protein or protein-nucleic acids interactions are a primary component, propose to purchase a spectropolarimeter capable of steady-state and stopped-flow circular dichroism (CD) studies. The instrument, manufactured by Applied Photophysics, is intended as a replacement and upgrade for an existing and outdated Jasco CD purchased in 1989. The existing instrument remains the only CD instrument on the NCSU campus and is used heavily by several faculty across the campus. However, it becomes increasingly difficult to maintain the instrument in working condition in order to acquire adequate data. The existing instrument will not provide reliable data below 200 nm, the temperature control is poor, the software and computer are antiquated, and it does not have stopped-flow or titration capabilities. In the past three years, the instrument has required several thousand dollars in repair costs to remain functional. The proposed instrument will enable the study of protein and nucleic acid conformations and provide information on protein and/or nucleic acid stability as well as protein folding mechanisms. Measurements of binding profiles will allow the determination of binding constants and stoichiometries, thermal dependence to circular dichroism parameters will be obtained, and kinetic reactions will be measured. The instrument will facilitate the current experimental approaches of the investigators which include NMR spectroscopy, fluorescence spectroscopy, X-ray crystallography, differential scanning and isothermal titration calorimetry, circular dichroism, mass spectrometry, and other biochemical and biophysical techniques. The steady-state and stopped-flow spectropolarimeter will complement NIH funded structural and functional studies focused on understanding the folding and assembly of caspases, transition state regulator proteins involved in bacterial response and protection, and characterization of modified nucleoside contributions to RNA and to RNA:peptide and RNA:metal interactions.

描述（由申请人提供）：三位在田间工作的NCSU科学家，其中蛋白质 - 蛋白质或蛋白核酸相互作用是主要成分，建议购买能够稳态和停止流量循环二色性（CD）研究的光谱极性计（CD）研究。该仪器是由Applied Pothysics制造的，旨在作为1989年购买的现有和过时的Jasco CD的替代品和升级。现有的仪器仍然是NCSU校园上唯一的CD仪器，并被校园中的几位教师大量使用。但是，在工作条件下保持仪器以获取足够的数据变得越来越困难。现有仪器将无法在200 nm以下提供可靠的数据，温度控制很差，软件和计算机已过时，并且没有停止流量或滴定功能。在过去的三年中，该工具需要数千美元的维修费用才能保持功能。所提出的仪器将能够研究蛋白质和核酸构象，并提供有关蛋白质和/或核酸稳定性以及蛋白质折叠机制的信息。结合曲线的测量将允许确定结合常数和石化图，将获得对圆二色性参数的热依赖性，并将测量动力学反应。该仪器将促进研究人员的当前实验方法，包括NMR光谱，荧光光谱，X射线晶体学，差异扫描和等温滴定热量法，圆形二色性，质谱法以及其他生物化学和生物学和生物物理技术。稳态和停止的光谱光谱仪将补充NIH资助的结构和功能研究，重点是理解胱天蛋白酶的折叠和组装，参与细菌反应和保护的过渡状态调节蛋白，以及对RNA和RNA的改性核苷贡献的表征，对RNA和RNA：肽和RNA：金属相互作用。