SC COBRE in Oxidants, Redox Balance, and Stress Signaling
Mass Spectrometry Core
This Core is housed within the MUSC Mass Spectrometry Facility which provides expertise, services, education, and training to enhance biomedical research endeavors through mass spectrometry-based proteomics. Currently there are over 50 investigators which utilize the facility for protein identification and characterization. Protein analysis includes in-gel or in-solution protease digestion, chromatographic separation and tandem mass spectrometric analysis of the resulting peptides, and interpretation of MS/MS data using Sequest or Mascot software. The facility also assists in the development of customized applications for the isolation, detection and characterization of posttranslationally modified peptides (e.g. phosphorylation, glycosylation, oxidation, glutathionylation, and O-GlcNAc modification). With the recent acquisition of the Orbitrap Elite Mass Spectrometer we are expanding our services to couple quantitative approaches (SILAC, iTRAQ, ICAT) to modification-specific experiments (eg., phosphoproteomics, redox proteomics). We are developing methodology to analyze alterations in posttranslational regulation that impact signal transduction, epigenetic modulation, and the response to therapeutics with the goal of enabling investigators to discover molecular mechanisms underlying disease progression and therapeutic responses that may not be revealed through genomic studies.
Instrumentation and associated proteomic applications available include:
Thermo LTQ XL Linear Ion Trap MS (2) (CID, PQD, ETD fragmentation); LC-MS/MS for protein identification and characterization of fragile modifications.
Applied Biosystems 4800 Plus MALDI-TOF-TOF Proteomics Analyzer; LC-MALDI-MS/MS for protein identification and quantitation of differentially expressed protein using iTRAQ reagents.
Bruker Autoflex III MALDI-TOF-TOF MS; MALDI Tissue Imaging.
Bruker Autoflex III MALDI-TOF MS; Molecular weight determination of intact proteins and peptides.
Bruker Solarix 7T Dual Source MALDI/ESI FT-ICR MS (CID and ECD Fragmentation); MALDI Tissue Imaging, Top-Down Protein Characterization
Thermo Orbitrap Elite with VelosPro Ion Trap MS (CID, HCD, ETD Fragmentation); LC-MS/MS for identification, characterization of modifications, quantitation of differential protein expression or posttranslational modification using SILAC or label free approaches, Top-Down Protein Characterization.
Associated HPLC systems (5 LC Packings nano-LC systems and 2 Dionex Probot MALDI Spotters for LC-MALDI). Agilent 3100 Off gel fractionator. Parker Nitrogen generator and Atlas Copco Air Compressor. MDS Arcturus XT Laser Capture Microdissection. Six Dell data workstations and a Dell Precision T5500 dual quad core workstation to support data interpretation. For archiving data, a Dell rack-mounted server with Quad Core Intel Xeon 3 GHz processors, 4 Gb RAM, and 1.5 TB RAID storage is monitored and automatically backed-up through the university archive system.
Additional proteomics software resources include:
Nonlinear Dynamics Progenesis SameSpots system. Genologics OMIX/Proteus system-for automated archiving as data is acquired. MUSC-developed software produced by John Schwacke. ProteoIQ. Mascot work station upgrade.
Lauren Ball, Ph.D.
Jennifer Bethard, M.S.
Research Associate of Cell and Molecular Pharmacology and Experimental Therapeutics