MRI: Acquisition of Advanced LC/MS/MS Instrumentation for Research and Teaching to Characterize Structure-Function Relationship Dynamics of Protein Systems

联邦资金将用于为北卡罗来纳州立大学(NCSU)分子和结构生物化学系的生物化学质谱学实验室购买先进的LC/MS/MS系统和辅助设备，用于研究蛋白质结构和蛋白质功能关系动力学。配置的仪器将用于确定各种生物系统中蛋白质的结构-功能方面，其中包括定量蛋白质组学分析，以确定翻译后修饰(PTM)以及在发现和假设驱动的研究中的蛋白质结合相互作用和化学计量学。分析测量将包括表征PTM，如磷酸化、乙酰化、糖基化和泛素化，以及这些修饰如何影响蛋白质的生化功能，包括修饰蛋白质与其底物结合的能力，与其他蛋白质相互作用，并促进进一步的PTM改变。购买的仪器将提供当前校内外任何LC/MS/MS仪器无法提供的独特功能，并将填补这一空白，以支持需要高级LC/MS/MS分析的不同大学的NC州立教师的研究，同时为学生和博士后提供直接培训，使他们能够成功完成他们的研究项目。该仪器将由19名教职员工的实验室使用，并将为大约4名高中生、38名本科生、34名研究生、13名博士后科学家和9名高级研究/访问科学家提供培训，预计将为NCSU的其他教职员工和学生提供随着他们研究的发展和需要进行LC/MS/MS分析蛋白质的途径。此外，这种仪器将用于特殊的研究实习项目，学生来自代表性不足的群体，还将用于生物化学专业的高级生物化学本科实验室课程(拥有近500个专业，这是全国第三大本科生物化学项目)，以鉴定和表征实验室中纯化的蛋白质。购买的LC/MS/MS系统将与核磁共振、X射线结晶学和电子冷冻显微镜一起作为研究蛋白质及其络合物结构和功能的工具，并将使NCSU的研究和教学项目能够更全面地培训学生和年轻研究人员生物化学、结构生物学和质谱分析，为成功的生物技术研究和教学生涯做好准备。除了在接入和能力方面显著改进NCSU的LC/MS/MS支持外，LC/MS/MS系统还将代表着在NCSU发展由生物核磁共振、X射线结晶学和质谱学组成的世界级结构生物化学系的长期目标的重大成就。2008年在波尔克霍尔建成了一个新的结构生物化学翼，其中包括核磁共振、X射线和质谱仪，并为这些领域的重大扩展提供了空间，证明了该大学对这一目标的承诺。收购这一用于研究蛋白质相互作用和结构-功能分析的高分辨率LC/MS/MS仪器，可以更充分地利用我们三个结构平台的组合优势。自从2007年我们购买了美国国家科学基金会资助的Q-TOF以来，我们目前唯一的高分辨率MS仪器，我们的成功已经导致校园对这种和更高分辨率MS功能的兴趣显著增加。新的LC/MS/MS系统将为本科生、研究生和博士后科学家提供前所未有的机会，为他们提供与技术相关的、全面的LC/MS/MS背景，这将使他们未来在其他教育项目或行业取得成功。有了这项技术，教师研究项目的进展将更快、更有效、更具科学影响力，这将使NCSU在资助和吸引顶尖人才教师方面在其他研究密集型大学中更具竞争力。由于PIs和其他用户对这项外展工作的强烈承诺，所请求的LC/MS/MS仪器和我们的MS设施作为一个整体将在部门和其他校园外展计划和活动中发挥积极作用，这些计划和活动旨在发展和培养初中和高中水平的早期对科学的兴趣，包括代表人数较少的群体(例如，ACS SEED和NSF AGEP计划)，最终目标是提高对生物科学的普遍认识，并鼓励有才华的年轻男性和女性选择研究和教学作为职业目标。

Federal funds are being used to purchase an advanced liquid chromatography-tandem mass spectrometry (LC/MS/MS) system and supporting equipment for the Biochemistry Mass Spectrometry Laboratory in the Department of Molecular and Structural Biochemistry at North Carolina State University (NCSU) for studying protein structure and protein functional relationship dynamics. The configured instrumentation will be used for ascertaining the structure-function aspects of proteins in a variety of biological systems, which includes quantitative proteomic analysis to determine post-translational modifications (PTMs) and protein binding interactions and stoichiometries in both discovery- and hypothesis-driven studies. Analytical measurements will include characterizing PTMs such as phosphorylation, acetylation, glycosylation, and ubiquitination, and how these modifications influence the biochemical functions of proteins, including the ability of modified proteins to bind to their substrates, interact with other proteins, and promote further PTM alterations. The purchased instrumentation will provide unique capabilities not possible with any current LC/MS/MS instrumentation on or off campus and will fill this void to support the research of NC State faculty in various colleges requiring advanced LC/MS/MS analysis while providing direct training of students and postdocs to enable successful completion of their research projects. The instrumentation will be used by the laboratories of 19 faculty members and will provide training for an estimated 4 high school students, 38 undergraduates, 34 graduate students, 13 postdoctoral scientists, and 9 senior research/visiting scientists with the anticipation of providing other NCSU faculty and students access as their research develops and requires LC/MS/MS analysis of proteins. In addition, this instrumentation will be used for special research internship programs with students from underrepresented groups and will also be used in the advanced biochemistry undergraduate laboratory course for biochemistry majors (with nearly 500 majors, this is the third largest undergraduate biochemistry program in the nation), to identify and characterize a protein purified in the laboratory. The purchased LC/MS/MS system will join NMR, X-ray crystallography, and electron cryo-microscopy as a tool for the study of the structure and function of proteins and their complexes and will enable the research and teaching programs at NCSU to more comprehensively train students and young investigators in biochemistry, structural biology, and mass spectrometry analysis to better prepare them for successful biotechnology research and teaching careers. In addition to providing significant improvements in LC/MS/MS support at NCSU, both in terms of access and capabilities, the LC/MS/MS system will represent a significant achievement in the long-range goal of developing a world class structural biochemistry department at NCSU comprised of biological NMR, X-ray crystallography, and mass spectrometry. The commitment of the University to this end is evidenced by the completion in 2008 of a new structural biochemistry wing in Polk Hall which houses NMR, and X-ray, and MS instrumentation and provides space for significant expansion in each of these areas. Acquisition of this high resolution LC/MS/MS instrumentation for studying protein interactions and structure-function analysis can be used to more fully leverage the combined strengths of our three structural platforms. Since the purchase of our NSF-funded Q-Tof in 2007, currently our only high resolution MS instrument, our successes have resulted in a significant increase in interest on campus for this and higher resolution MS capabilities. The new LC/MS/MS system will provide unprecedented access for undergraduates, graduate students, and postdoctoral scientists, providing them with a technologically relevant, comprehensive LC/MS/MS background which will enable their future success in other educational programs or in industry. With access to this technology, faculty research programs will be progressed more quickly, efficiently, and with greater scientific impact which will place NCSU more competitively among other research-intensive universities for funding and attracting top talent faculty. Because of the strong commitment of the PIs and other users on this proposal for outreach work, the requested LC/MS/MS instrument and our MS facility as a whole will play an active role in departmental and other campus outreach programs and activities aimed at developing and fostering early interest in science at the middle and high school levels, including underrepresented groups (for example, ACS SEED and NSF AGEP programs), with the end goals of raising general awareness of the biological sciences and encouraging talented young men and women to choose research and teaching as a career goal.