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Ashley Phetsanthad Thesis Defense
March 15 @ 3:00 pm - 4:00 pm
Ph.D. DISSERTATION DEFENSE
Ashley Phetsanthad
Developing Mass Spectrometry-Based Strategies for the Enhanced Characterization of Biomolecules in the Nervous System
Li Research Group
Wednesday, March 15th at 3:00 pm
Room 1335 of the Health Sciences Learning Center
Or virtually: https://uwmadison.zoom.us/j/97063598170?pwd=SHAzL1kwTmcyYm1PdkdzNVNLNUZ6dz09
Neuronal signaling is essential for the body to recognize and respond to stimuli, both internal and external. Neurons rely on proteins, neuropeptides, lipids, and other small molecules to properly receive and transmit signals; however, this can be affected or disrupted due to changes in structure, regulation, or the presence of any post-translational modifications (PTMs). Awareness of modification states is critical to understanding the molecular mechanisms involved with basic biological function, as well as those altered by various neurological diseases and disorders. While elucidating PTM states can be difficult due to heterogeneity and low stoichiometry, the sensitivity and rapid data acquisition capabilities of mass spectrometry (MS) provides the ability for quantitation and accurate PTM characterization.
I will demonstrate a workflow to select data-independent acquisition MS parameters systematically using definitive screening designs to improve neuropeptide identification. Neuropeptides will be further characterized using an improved workflow to greatly expand the library of known endogenous glycosylated neuropeptides in Callinectes sapidus neuronal tissues. I also describe a method to decrease charge state bias in structural characterization by ion mobility spectrometry MS and its utility in efforts to investigate the indirect effects of glycoprotein sialylation on neuronal cytotoxicity. In summary, this dissertation details the development of MS methodologies to improve our understanding of the endogenous peptides and proteins involved in neuronal function.