Defining the conformational control of nitric oxide synthases by a multipronged approach

通过多管齐下的方法定义一氧化氮合酶的构象控制

• 批准号: 10385652
• 负责人: Changjian Feng
• 金额: $ 10.5万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10385652
• 关键词: Administrative Supplement; Alzheimer' s Disease; Anabolism; Award; Bioinorganic Chemistry; Complex Mixtures; Development; Disease; Environment; Equipment; Family; Financial Support; Funding; Institution; Investments; Laboratories; Lead; Liquid Chromatography; Methods; Molecular Conformation; Multienzyme Complexes; New Mexico; Nitric Oxide; Nitric Oxide Synthase; Parents; Pharmacy facility; Process; Production; Productivity; Proteins; Research; Research Personnel; Site; Stroke; System; Techniques; United States National Institutes of Health; Universities; Work; biophysical chemistry; college; effective therapy; fast protein liquid chromatography; instrumentation; milligram; novel therapeutic intervention; novel therapeutics; parent project; programs; protein complex; protein purification; spectroscopic survey; unnatural amino acids

PROJECT SUMMARY The parent award is on deciphering conformational dynamics of nitric oxide synthase proteins through an integrated approach using advanced spectroscopic techniques. Detailed spectroscopic studies require milligram amounts of highly purified protein. Fast protein liquid chromatography (FPLC) is a form of liquid chromatography used to purify a target protein from a complex mixture. Our current FPLC has been a workhorse, but is near the end of operational lifetime. This administrative supplement is being requested to allow acquisition of a state-of-the-art ÄKTA pure 25 system. It has direct relevance to and immediate utility in enhancing our laboratory’s protein purification capabilities. As such, this equipment will greatly enhance our research productivity for the duration of the project, and will help us maintain a strong research program. Importantly, the unique capabilities of the ÄKTA pure 25 system will be indispensable for our efforts to develop and optimize methods for site-specifically incorporating unnatural amino acids into large, complex proteins with high fidelity, yield, and purity. Our findings will therefore be of great utility for other investigators using this process for other large, complex enzymes. Therefore, the requested equipment will help the parent project to exert a sustained, powerful influence on the research fields of bioinorganic and biophysical chemistry. Moreover, the administrative supplement will enhance the scientific environment for NIH-funded investigators at the University of New Mexico, both within and outside of the College of Pharmacy. To our knowledge, no comparable instrumentation exists in educational institutions within the region. Dean Godwin has committed substantial financial support to this supplement request since this is a great investment for the College. Taken as a whole, the proposed supplement will increase the parent award’s overall impact within the original scope of award, as well as bolster research quality and depth at our institution.

项目摘要 父母奖是通过一个解释一氧化氮合酶蛋白质构象动力学的 采用先进的光谱技术的综合方法。详细的光谱研究需要 毫克量的高度纯化的蛋白质。快速蛋白质液相色谱法（FPLC）是一种液相色谱法， 用于从复杂混合物中纯化靶蛋白的层析。我们目前的FPLC一直是一个 主力，但接近年底的运作寿命。要求提供这一行政补充材料是为了 允许获得最先进的ÄKTA pure 25系统。它与以下方面有直接的关系和直接的效用： 增强我们实验室的蛋白质纯化能力因此，该设备将大大提高我们的 研究生产力的项目期间，并将帮助我们保持一个强大的研究计划。 重要的是，<$KTA pure 25系统的独特功能对于我们开发 并优化了将非天然氨基酸位点特异性地掺入大型复杂蛋白质的方法， 高保真度、产率和纯度。因此，我们的研究结果将是非常有用的其他调查人员使用这一点， 其他大型复杂酶的过程。因此，所要求的设备将有助于父项目 对生物无机化学和生物物理化学的研究领域产生了持续的，强大的影响。 此外，行政补充将改善NIH资助的研究人员的科学环境 在新墨西哥州的大学，无论是在药学院内外。据我们所知，没有 该区域的教育机构也有类似的工具。迪恩·戈德温承诺 这一补充请求得到了大量的财政支持，因为这对学院来说是一项巨大的投资。采取 总的来说，建议的补助金将在原有范围内增加家长奖的整体影响 奖励，以及加强研究质量和深度在我们的机构。