Acquisition of Trapped Ion Mobility Spectrometry Time of Flight Mass Spectrometer - PROJECT SUMMARY This application requests funds to acquire a trapped ion mobility time of flight (timsTOF) mass spectrometer (MS) from Bruker with online PASEF (Parallel Accumulation Serial Fragmentation) technology to be deployed at the Purdue Proteomics Facility. The application includes project descriptions from seven major and seven minor users that will benefit from the access to this instrument. The timsTOF Pro offers rapid, high-resolution separation of peptides and proteins based on structure (ion conformation) and mass-to-charge (m/z) ratios. The unique feature of dual-TIMS funnel provides enhanced ability to separate ions by differences in conformation, thus allowing separation of isobaric and isomeric species, such as phosphopeptides and glycopeptides positional isomers, which are not easily resolved by m/z and LC retention time separation. The novel TIMScore algorithm with PaSER (Parallel database Search Engine in Real-time) technology distinguishes the timsTOF Pro from other commercial IMS instruments. An experienced facility director, dedicated staffs, bioinformatics support, internal advisory committee comprising leading experts in mass spectrometry, strong institutional commitment and responsible operation and management plans are described. The Office of the Executive Vice President for Research and Partnership and the Bindley Bioscience Center will provide excellent infrastructure and commitment to additional funds toward the extended service contract to insulate users from increased fees. Purdue will also provide partial funding over the first five years for a staff scientist who will be hired to support the instrument operation and maintenance. Of the 4,000 hours of estimated accessible user time (AUT) of the instrument, seventy-five percent is allocated to NIH-supported research projects, and remaining 25% will be used by other core users and for cleaning, calibration, annual PM, method development, and user training. Projects described in this application aim to address changes in protein modifications, and protein-protein and protein-ligand interactions, critical in cellular signaling and disease pathologies. Thus, access to the timsTOF Pro MS will benefit many NIH supported projects that focus in signal transduction, therapeutic protein characterization, structural biology, biomarker discovery, and infectious diseases.
