MRI: Track #1: Acquisition of a MALDI-TOF-TOF Mass Spectrometer for Graduate and Undergraduate Research and Education

This award is funded by the Major Research Instrumentation Program and the Chemistry Research Instrumentation Program. Professor Andreas Franz from the University of the Pacific (Stockton, California), on behalf of 9 investigators in 4 departments across the university, is acquiring a matrix-assisted laser desorption / ionization time-of-flight mass spectrometer (MALDI-TOF-MS). Mass spectrometry (MS) can detect very small amounts of substances by ionization and is a central analytical method in science. MALDI-TOF-MS uses laser pulses to produce a wide range of ions, especially from high molecular weight biomolecules such as peptides, proteins, oligonucleotides, carbohydrates, and synthetic polymers. The TOF detector is akin to a racetrack where ions are separated by mass-to-charge ratio on their way to the finish line (detector). High-speed electronics ("photo-finish") can differentiate very small differences in arrival time and, therefore, mass. The acquisition strengthens collaborative research infrastructure at the university and broadens participation by giving hands-on access to a diverse student population. To advance the university's outreach activities and improve educational opportunities, regional research groups at other universities and junior colleges will use the instrument through remote-access platforms as well as through on-campus visits and workshops. With the new instrument structural studies of biomacromolecules and polymers will be strengthened in general, and many exciting projects are enabled. We will advance knowledge in the following areas: a) glycan abundance and microheterogeneity of protein glycosylation as it relates to protein stability and biochemical function; b) use of yeast as high-quality/high-capacity protein expression system; c) DNA quadruplex stability as it relates to ageing; d) cytokine inhibitors for the treatment of heart diseases; e) rapid in-situ identification of microbes and microbial cell-cell communication; f) inhibition of SARS-CoV-2. Oligosaccharides have many complex functions in biochemistry, and great progress has been made to unravel their roles in many central life processes related to health and disease. Unambiguous structure elucidation is critical before quantification and correlation to disease is possible. The UV-laser light source of the instrument will generate high-energy ions from oligosaccharides with a covalent photo-sensitive tag. The photosensitive tag causes more extensive fragmentation than usual for rich structure information. The method will be applied to study glycosylation aberrations in proteins expressed in yeast and thus, shed light on the biochemical machinery of the yeast cell. Another project will generate insight into DNA quadruplex stability and advance the discovery of covalent adducts between DNA and small molecule therapeutics that induce quadruplex formation to combat cancer. In another project the instrument will be central to screening proteins, peptides. The focus is on cancer-associated proteins that are thought to be "undruggable". New strategies for targeting these proteins will be explored in which MALDI-TOF-MS serves the purpose of structure confirmation of point-mutated protein variants and the possible discovery of strongly binding non-covalent ligands by high-throughput methods. Similar strategies based on ion-intensity fading in MALDI-TOF-MS will be used to discover peptides that bind strongly to viral proteins, specifically those that mediate binding of the SARS-CoV-2 virus. Lastly, MALDI-TOF-MS will help with the high-throughput analysis of complex natural products from marine microorganisms and provide information on how microorganisms interact and communicate with each other. For all researchers, the instrument's automation platform will increase sample throughput for quick lead identification; will make possible the accelerated discovery of new therapeutics against human diseases; will aid in the discovery of therapeutics against age-related diseases. Faster acquisition of spectra will free up time to make the instrument accessible to nearby institutions and include MALDI-TOF-MS into the academic curriculum for young aspiring scientists. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.