Acquisition of Transient Kinetic Equipment

瞬态动力学设备的购置

• 批准号: 9512166
• 负责人: Michael Schimerlik
• 金额: $ 13.84万
• 依托单位: Oregon State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9512166&HistoricalAwards=false
• 关键词: Acquisition; Transient; Kinetic; Equipment

Funds are requested to purchase transient kinetic equipment to be employed in research projects from eleven different laboratories at C)region State University. The users represent four departments: Biochemistry and Biophysics, Agricultural Chemistry, the College of Pharmacy and the Department of Food Science and Technology. The equipment will allow the users to pursue new approaches to their respective research problems in the areas of protein folding, protein-protein and protein-DNA interactions, DNA and protein structure, protein- and receptorligand interactions, kinetic studies of enzymes, the coupling of receptors to ion channels and the relationship between mitochondrial integrity, oxidative stress and Ca2+ overload. The availability of this equipment should serve to attract other researchers at Oregon State University interested in mechanistic studies and will therefore serve as an important resource for faculty, postdoctorals, graduate students and undergraduates pursuing these research areas. Since the equipment is novel to Oregon, it may also attract users from other institutions within the state. One major feature of the instrumentation is the ease of data collection and analysis. This should facilitate user training and increase the enthusiasm of users in applying kinetic, mechanistic approaches in their research. The principal investigator is Dr. Michael Schimerlik. The PI received his Ph.D. in enzyme kinetics with Professor W.W. Cleland at Wisconsin and training in transient kinetics while a postdoctoral at The California Institute of Technology with Professor M.A. Raftery where he conducted stopped-flow studies of ligand interactions with the nicotinic acetylcholine receptor. Since coming to Oregon State, the PI has published manuscripts concerning transient kinetic studies in the areas of protein-protein interactions and enzyme mechanisms and is capable of analysis of kinetic mechanisms by fitting data to analytically derived rate equations or, i n more complicated instances, using computer simulations to examine mechanistic models. Several other investigators (Drs. Penner, van Holde, Ho, Anderson and Hsu) also possess expertise in transient kinetic theory. Transient kinetic equipment existing at Oregon State is about fifteen years old, and no longer supported by the manufacturer. No facilities are available for quench flow experiments for use in combination with state of the art facilities at the institution in mass spectrometry or NMR for studies of protein folding. No facilities are available for rapid acquisition of circular dichroism spectra to examine structural changes that occur in protein folding, ligand binding, DNA and protein structural transitions, or macromolecular assembly or dissociation reactions. The abilities to acquire fluorescence or absorbance spectra as a function of time, or to measure time-dependent changes in fluorescence polarization are also lacking. These techniques provide additional sensitivity and enhance the analysis of complicated mechanisms. Most importantly, the software for data acquisition and analysis is far superior to that currently available. In summary, the proposed kinetic equipment will add important new capabilities in transient kinetics not currently available at Oregon State University. In addition it will provide greatly enhanced capabilities over that of our currently available, outdated equipment both in the type of data acquired and in its analysis. This will not only expand the research horizons of laboratories doing kinetic studies but, also because of its "user-friendliness", serve to attract postdoctorals, graduate students and undergraduates so that these individuals will receive training in transient kinetic methods.

申请资金用于购买瞬态动力学设备，用于C)地区州立大学11个不同实验室的研究项目。使用者代表四个系：生物化学与生物物理系、农业化学系、药学院和食品科学与技术系。该设备将允许用户在蛋白质折叠、蛋白质-蛋白质和蛋白质-DNA相互作用、DNA和蛋白质结构、蛋白质和受体配体相互作用、酶的动力学研究、受体与离子通道的偶联以及线粒体完整性、氧化应激和Ca2+过载之间的关系等领域寻求新的研究方法。该设备的可用性将有助于吸引俄勒冈州立大学对机械研究感兴趣的其他研究人员，因此将成为从事这些研究领域的教师、博士后、研究生和本科生的重要资源。由于这种设备对俄勒冈州来说是新奇的，它也可能吸引州内其他机构的用户。该仪器的一个主要特点是易于收集和分析数据。这将有助于用户培训和提高用户在研究中应用动力学和机械方法的热情。首席研究员是Michael Schimerlik博士。他在威斯康星大学获得酶动力学博士学位，在加州理工学院获得瞬态动力学博士学位，在加州理工学院获得M.A. Raftery教授的博士后学位，在那里他进行了配体与烟碱乙酰胆碱受体相互作用的停流研究。自从来到俄勒冈州以来，PI已经发表了关于蛋白质-蛋白质相互作用和酶机制领域的瞬态动力学研究的手稿，并且能够通过拟合数据来分析导出的速率方程来分析动力学机制，或者在更复杂的情况下，使用计算机模拟来检查机制模型。其他几位研究者(dr。Penner, van Holde, Ho， Anderson和Hsu)也拥有瞬态动力学理论的专业知识。俄勒冈州现有的瞬态动力设备大约有15年的历史，不再由制造商提供支持。没有可用于淬火流动实验的设备，无法与该机构最先进的质谱或核磁共振设备相结合，用于蛋白质折叠的研究。没有设备可用于快速获取圆二色光谱，以检查发生在蛋白质折叠，配体结合，DNA和蛋白质结构转变，或大分子组装或解离反应中的结构变化。获取荧光或吸光度光谱作为时间函数的能力，或测量荧光偏振随时间变化的能力也缺乏。这些技术提供了额外的灵敏度，并加强了对复杂机构的分析。最重要的是，用于数据采集和分析的软件远远优于目前可用的软件。总之，拟议的动力学设备将为瞬态动力学增加重要的新能力，目前在俄勒冈州立大学还没有。此外，在获取数据的类型和分析方面，它将大大增强我们目前可用的过时设备的能力。这不仅将扩大动力学研究实验室的研究视野，而且由于其“用户友好”，有助于吸引博士后、研究生和本科生，以便这些人接受瞬态动力学方法的培训。