MODEL PROTEIN PRODUCTION FOR TIME-RESOLVED 2D-ELDOR, PDS, AND MQC ESR

时间分辨 2D-ELDOR、PDS 和 MQC ESR 的蛋白质生产模型

• 批准号: 8364074
• 负责人: ELKA R GEORGIEVA
• 金额: $ 0.67万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364074
• 关键词: Biological Models; Calmodulin; Collaborations; Coupled; DNA; Development; Funding; Grant; Housing; Methods; Modeling; Muramidase; National Center for Research Resources; Principal Investigator; Production; Proteins; Research; Research Infrastructure; Resources; Sampling; Source; Spectrum Analysis; Spin Labels; Technology; Time; United States National Institutes of Health; cost; interest

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Good model systems are needed to develop methods, which is certainly true of PDS. Long organic biradicals, polyradicals, and supramolecular assemblies were instrumental for development of DEER. However they are difficult to synthesize and require expertise of a good organic chemist, who should be interested in such collaborations. In our development of DQC method we could not rely on this but made a good use of few available biradicals with distances in the range of 10-30 ¿. But when we needed a longer distances, we had to reach to Gunnar Jeschke for the sample. However we did find that for example DNA duplexes or some proteins can also contribute good model systems and are relatively easy to produce in house in required quantities. For the project development on time-resolved 2D-ELDOR spectroscopy, coupled with stopped flow or continuous rapid flow, we require large amounts (up to grams) of spin-labeled model proteins. For example T4 Lysozyme and Calmodulin are relatively small and stable proteins that can be expressed in required quantities with a moderate effort. In general proteins and DNA could provide us with all required distances and a variety of spin configurations. Furthermore, they are much more relevant to our research.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供 该子项目的主要支持。 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源的子项目可能列出的总成本。 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 开发方法需要良好的模型系统，长有机双自由基、多自由基和超分子组装体对于 DEER 的开发至关重要，但它们很难合成，需要优秀的有机化学家的专业知识。在我们开发 DQC 方法时，我们不能依赖于此，而是充分利用了距离在 10-30 范围内的少数可用双自由基。但当我们需要更远的距离时，我们必须联系 Gunnar Jeschke 获取样品。然而，我们确实发现，例如 DNA 双链体或某些蛋白质也可以提供良好的模型系统，并且相对容易在内部生产所需的数量。时间分辨 2D-ELDOR 光谱的项目开发，加上停止流动或连续快速流动，我们需要大量（最多克）的自旋标记模型蛋白，例如 T4 溶菌酶和钙调蛋白。是相对较小且稳定的蛋白质，可以通过适度的努力以所需的量表达。一般来说，蛋白质和 DNA 可以为我们提供所有所需的距离和各种旋转构型。此外，它们与我们的研究更相关。