Post-translational modifications (PTMs) are chemical changes that proteins undergo after they are synthesized in cells. These modifications are incredibly diverse, including the addition of functional groups, cleavage of peptide bonds, or structural changes, such as phosphorylation, methylation, acetylation, ubiquitination, etc. PTMs can impact almost every aspect of a protein's life cycle, including its folding, stability, activity, interactions, and localization within the cell. They play a pivotal role in rapid regulation of cellular processes and signaling pathways, which are essential for maintaining cellular homeostasis and responding to external stimuli. Aberrations in PTMs are linked to various diseases, including cancer, neurodegenerative disorders, and metabolic syndromes, highlighting their significance in health and disease. Therefore, sensitive methods for the characterization of PTMs is crucial to understand cellular processes and disease mechanisms in detail. Moreover, PTMs offer potential targets for therapeutic intervention, as modulating these modifications can alter disease progression. Finally, research in PTMs contributes to the development of biomarkers for disease diagnosis and prognosis, further underscoring their importance in biomedical science.
We have previously developed several analytical methods for characterization of PTMs using biological mass spectrometry (e.g., 1-4). In the present presentation we will present these highly optimized methods in a comprehensive workflow content allowing quantitative assessment of multiple PTMs from the same set of samples. The workflow includes assessing phosphorylation, lysine acetylation, various cysteine modifications, glycosylation and lipidation. We show that such a comprehensive workflow significantly increases the coverage of the dynamic proteome in cells, tissues, and liquid biopsies. We will show examples of the application of such a workflow on studying cellular signaling in various cells such as T-cells and neurons, including neurons in human brain organoids.
Literature:
[1] Larsen MR et al., MCP 2005, 4(7), 873-86. [2] Larsen, MR., et al., MCP. 2007, 6(10), 1778-1787. [3] Huang H et al., J Proteomics. 2023, 273:104796. [4] Huang H et al MCP 2016, 15(10):3282-3296.
