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

联邦资金正用于为北卡罗来纳州州立大学分子和结构生物化学系生物化学质谱实验室购买先进的液相色谱-串联质谱（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蛋白质在各种生物系统中的结构-功能方面，其中包括定量蛋白质组学分析，以确定发现和假设驱动研究中的翻译后修饰（PTM）和蛋白质结合相互作用和化学计量。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，与其他蛋白质相互作用，并促进进一步PTM改变的能力。购买的仪器将提供独特的能力不可能与任何当前的LC/MS/MS仪器或校外，并将填补这一空白，以支持NC国家教师在各个学院的研究需要先进的LC/MS/MS分析，同时提供学生和博士后的直接培训，使他们的研究项目的成功完成。该仪器将由19名教师的实验室使用，并将为估计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，预计将为其他NCSU教师和学生提供访问，因为他们的研究发展需要LC/MS/MS分析蛋白质。此外，这种仪器将用于特殊的研究实习计划，学生来自代表性不足的群体，也将用于高级生物化学本科实验室课程的生物化学专业（有近500个专业，这是第三大本科生物化学计划在全国），以确定和表征蛋白质纯化在实验室。购买的LC/MS/MS系统将加入NMR，X射线晶体学和电子冷冻显微镜作为研究蛋白质及其复合物的结构和功能的工具，并将使NCSU的研究和教学计划能够更全面地培训学生和年轻的研究人员在生物化学，结构生物学和质谱分析，以更好地为成功的生物技术研究和教学事业做好准备。除了在NCSU提供LC/MS/MS支持方面的显着改进外，在访问和能力方面，LC/MS/MS系统将代表在NCSU开发世界级结构生物化学部门的长期目标方面的重大成就。该大学的承诺，以实现这一目标是在2008年完成了一个新的结构生物化学翼波尔克大厅，房子核磁共振，X射线和MS仪器，并提供空间，在每个这些领域的显着扩展。获得这种高分辨率LC/MS/MS仪器用于研究蛋白质相互作用和结构-功能分析，可用于更充分地利用我们三个结构平台的综合优势。自从2007年购买了我们NSF资助的Q-Tof（目前我们唯一的高分辨率MS仪器）以来，我们的成功使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，为他们提供技术相关，全面的LC/MS/MS背景，这将使他们未来在其他教育计划或行业中取得成功。有了这项技术，教师的研究计划将更快，更有效地进行，并具有更大的科学影响力，这将使NCSU在其他研究密集型大学中更具竞争力，以获得资金和吸引顶尖人才。由于PI和其他用户对这项推广工作提案的坚定承诺，所要求的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推广计划和活动中发挥积极作用，旨在培养和培养初中和高中阶段对科学的早期兴趣，包括代表性不足的群体（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女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.