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

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.