University of Victoria
Knowledge of biochemical pathways is part of the foundation of basic and applied research in the field of biochemistry. Mass spectrometers are indispensable tools for the study of biomolecules due to the afforded specificity, multiplexing and quantitative capabilities for the analysis of a broad range of analytes, encompassing small molecules such as metabolites, and multicomponent macro-molecules, such as protein complexes. High mass accuracy and high resolving power are the two main factors that make mass spectrometers inherently selective, and when combined with an internal standard spike in, absolute specificity is achievable. Additionally, mass spectrometers can be utilized to scan mass to charge ratios on the order of milliseconds which enhances multiplexing. When coupled with upstream separation strategies, such as liquid chromatography, hundreds to thousands of compounds can be identified and/or quantified within a matter of hours.
Analyte quantification can be performed in relative terms, for example comparing healthy to disease subjects, and/or accurate concentrations can be measured using spiked in standards. Other proteomics specific applications of mass spectrometry include enzymatic treatment of samples and subsequent identification and quantification of site-specific post-translational modifications. Lastly, protein-protein interactions and protein complexes can be characterized using cross-linking strategies and structural proteomics performed utilizing deuterium exchange methodologies. Each of the aforementioned applications requires unique operational and instrumental setups and this collection aims to facilitate the dissemination of these methodologies to other researchers.