PROJECT SUMMARY The Advanced Science Research Center (ASRC) of the City University of New York (CUNY) is requesting funds from NIH through the High-End Instrumentation program to purchase a timsTOF-FleX MALDI-2, high- resolution mass-spectrometer with trapped-ion mobility (TIMS) technology and a dual ion-source platform for metabolomics and proteomics studies. This instrument will support biomedical research projects of twelve laboratories from within CUNY and nearby institutions, including investigators from the Structural Biology and Neuroscience Initiatives at the ASRC, City College of New York, neighboring institutions in New York (Columbia University, Rockefeller University), and other laboratories in the region and nationally. The projects span biochemistry and structural biology from macromolecular to metabolomic levels, with two general areas of focus: (1) conformational dynamics of light-sensitive proteins, kinases, large systems as platelet integrins, and ryanodine receptors; (2) metabolic signals regulating tumor progression, and axonal regeneration and to metabolic control of brain cell population dynamics, with an emphasis on epigenetic regulation of glial cell differentiation and dietary effects on metabolites affecting neurodegeneration. The anticipated outcomes are better mechanistic definitions of processes regulating critical biological functions, giving insights into fundamental biology, ways that these mechanisms are impacted by disease, and routes to successful therapeutic and biotechnology strategies. Currently, no similar instruments are available in shared facilities in the New York City area. The novel technology incorporated into the proposed timsTOF-FleX MALDI-2 instrument will provide us with tools to significantly enhance the sensitivity and resolution of our mass measurements, enabling separation of challenging isomeric and isobaric species, confident detection and identification of many more molecular compounds in various Omics studies, and use of Hydrogen-Deuterium Exchange mass-spectrometry to increasingly complex macromolecular assemblies. Finally, the dual MALDI-MS and ESI-MS platform of timsTOF-fleX MALDI-2 will provide our users with the ability to map molecular distributions in tissue in situ, identify regions of interest that express the desired molecular profile, and selectively target these subpopulations for integrated Omics analyses. Thus, should this proposal be approved by NIH, the novel instrument will advance a number of collaborative research efforts in biomedical research being conducted at the ASRC and beyond, expand training opportunities for early career scientists. The instrument will be housed in the ASRC building and managed by the PI within the ASRC Mass- Spectrometry Core Shared Facility. All projects that require integrated MALDI-Imaging and complementary traditional Omics analysis will be supported by the MALDI-MS Imaging Joint Facility, co-directed by the PI and Dr. Ye He, director of the ASRC Live Imaging and Bioenergetics Facility.

PROJECT NARRATIVE This instrument will support biomedical research projects of twelve laboratories from within CUNY and nearby institutions, including investigators from the Structural Biology and Neuroscience Initiatives at the ASRC, neighboring institutions in New York, and other laboratories located throughout the region and nation. The anticipated outcomes are improved instrumental sensitivity, resolution, and other technical capabilities that will enable a diverse range of currently inaccessible experiments into fundamental mechanisms of biological control, critical for understanding life and disease at the molecular and level with implications for therapeutic and biotechnology applications.