Facility Details

The goal of the SC COBRE in Oxidants, Redox Balance and Signaling Proteomics Core is to provide state-of-the-art LC-MS/MS instrumentation, expertise, and training for comprehensive proteomic analyses to advance the research endeavors of investigators with interests in redox signaling. Dedicated core personnel assist with experimental design, method development, data acquisition, and computational analysis for protein characterization, interactions, and quantitative proteomic applications using metabolic labeling (SILAC), isobaric tandem mass tagging (TMT), and label free quantitation (MaxQuant LFQ). The core utilizes complementary peptide fragmentation modes (CID, HCD, ETD, EThcD, UVPD) as needed to characterize post-translational modifications of cysteine (sulfenic, sulfinic, sulfonic acid, S-glutathionylation, disulfide bonds); arginine (glycation by methylglyoxal, dihydroxyimidazolidine); tyrosine (nitrated and crosslinked), serine (O-GlcNAc and phosphorylation); lysine (acetylation, ubiquitylation, glycation, hydroxylation); and N- and O-linked glycosylation.

COBRE investigators have sequenced putative biomarker peptides discovered by MALDI-tissue imaging mass spectrometry and identified differentially expressed or post-translationally regulated proteins following genetic manipulation of anti-oxidant enzymes, drug treatment, or disease. Methodology has been established to determine the effects of altered redox-balance on differential protein abundance in FAC-sorted cell populations, primary cells, and exosomes. COBRE investigators have also utilized quantitative proteomics to elucidate the mechanism of drug action and identify the targets of drugs identified in phenotypic screens. We will continue to assist COBRE investigators and members of the COBRE Redox Center with customized method development as needed to advance their research endeavors.

Instrumentation available for quantitative LC-MS/MS-based proteomics include a ThermoScientific Orbitrap Fusion Lumos ETD/UVPD MS with EASY nanoUHPLC 1300 and an Orbitrap Elite ETD MS with Ultimate 3000 nanoLC acquired through the NIH Shared Instrumentation Program (S10 OD010731 and S10 OD025126, PI Ball). The core recently acquired a Waters Xevo TQ-S triple quadrupole MS with M class LC which will expand our capabilities to include absolute quantitation and multiple reaction monitoring (MRM) assays for targeted proteomics. Instrumentation is housed within the University supported MUSC Mass Spectrometry Facility and Proteomics Center.