Proteomics Core

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Proteomics Core – Project Summary The broad objective of the Proteomics Core in the SC COBRE in Oxidants, Redox Balance, and Stress Signaling (Redox COBRE) is to provide state-of-the-art, mass spectrometry (MS)-based proteomic capabilities to enable elucidation of redox signaling mechanisms underlying disease, therapeutic efficacy of cells and drugs, and therapeutic resistance. The Core provides expertise, technical assistance, instrumentation, and interpretation of analytical results for quantitative proteomic experiments and the characterization of redox-sensitive post- translational modifications. During Phase I and II, acquisition of the Orbitrap Elite ETD MS and Orbitrap Fusion Lumos ETD/UVPD MS nano-LC-MS/MS systems (NIH S10 OD010731 and S10 OD025126, PI: Lauren Ball) permitted detection of challenging modifications using complementary peptide fragmentation approaches (ETD and EThcD) and the development of advanced, quantitative proteomic workflows as needed by COBRE investigators. During Phase III, the Core will further develop and update these capabilities to reflect rapid advances in instrumentation, software, and methodologies relevant to redox proteomics. Additionally, in response to the needs of COBRE-affiliated investigators, proteomic tissue imaging capabilities using MALDI- Imaging MS (IMS) will be provided to discern the spatial distribution of enzymatically-digested peptides within tissue biospecimens on slides for the discovery of putative biomarkers and mechanisms underlying disease progression. Detection of disease-specific proteins and redox-sensitive post-translational modifications by LC- MS/MS and IMS has raised the need for targeted proteomic capabilities to verify these findings and further advance translational studies. Providing this complement of proteomic technologies will build on the successes of the Core and position the Core to establish a comprehensive, independent resource. The Core has leveraged previous investments made by the NIGMS during Phase I and II of the COBRE program, the OD and NIGMS for Shared Instrumentation Grants, the NCI Cancer Center Support Grant, and long-standing, strong institutional commitment to develop into a successful institutional resource critical to the needs of Redox COBRE investigators. At present there are 19 COBRE-affiliated users of the Proteomics Core that are PIs of 15 active NIH awards that rely on the LC-MS/MS capabilities, five of these awards also depend on IMS capabilities.