Bottom-Up Proteomic Techniques to Interrogate the Mycobacterial Proteome

Tuberculosis (TB) is a respiratory disease caused by the pathogen Mycobacterium tuberculosis (M. tb). This disease is the largest global cause of death by infectious disease, causing illness in 10.8 million people and 1.25 million deaths annually. Despite its major impact on human health, prevention and treatment measures to this disease are limited. Antibiotic treatments are reducing in efficacy due to the onset of multidrug-resistant tuberculosis (MDR-TB). Several techniques are used to study pathogenic mycobacteria. One of them is bottom-up proteomics, an analytical technique used to identify and quantify proteins in biological systems. While proteomics is ubiquitous today, it is often used as a black-box technique when applied to the life sciences. Innovations to proteomics in the analytical community rarely translate into new fit-for-purpose approaches to study disease and pathogens. The result is a gap in knowledge between the most up-to-date analytical methods and the forefront of mycobacterial research. In my dissertation, I describe the contributions I have made to bridge the gap between the mycobacterial research community and the bottom-up proteomics community. I describe two projects in which I apply custom data analysis techniques to my proteomic data in order to discover novel biological conclusions. In another project, I describe a new sample preparation technique I created specifically to quantify mycobacterial protein N-terminal acetylation, a ubiquitous protein post-translational modification. In my final chapter, I detail an aspect of my graduate career outside of bioanalytical chemistry—leadership in laboratory safety. I describe the history of my work as a leader in my department’s Laboratory Safety Officer Program, as well as the later contributions I made in organizing a 2-day Summit sponsored by the American Chemical Society.