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Proteomics Core
Proteomics Core is the central resource for mass spectrometry based proteomics within the NHLBI. The Core staff help collaborators design proteomics experiments in a way that are compatible with mass spectrometry and to modify protocols if needed to ensure they will provide high quality data. An important aspect of the Core’s mission is to teach users new methods, techniques and instruments. The Core also continually innovates on existing technologies.
Our workflows for relative protein quantitation are based on DIGE, label-free and iTRAQ approaches. We can also help investigators identify and quantify protein posttranslational modifications, including phosphorylation, nitrosylation, acetylation, ubiquitination, etc. We provide training in proper sample preparation and lead the researchers through mass spectrometric analysis to data searching and interpretation. Users have access to a variety of proteomics software platforms (Mascot, Sequest, Proteome Discoverer, Scaffold, Protein Pilot) for re-searching the data or viewing the results.
In addition to helping the NHLBI investigators, we develop new approaches for characterization of posttranslational modifications and absolute protein quantitation.
The NHLBI Proteomics Core was a recipient of the 2012 NHLBI Orloff Award for outstanding research support.
Recent studies and method development include:
- Acetylation on lysine (developing workflows for identification and occupancy measurements)
- Nitrosylation on cysteine (developing workflows for identification and occupancy measurements)
- Ubiquitination and SUMOylation (workflows for identification and quantitation)
- Development of ‘mito-panel’ – using targeted analysis on Velos to follow specific mitochondrial proteins
- Depletion of the most abundant serum proteins to study the protein differences in patients’ samples
- Absolute protein quantitation using SRM on Velos instruments
- Development of iTRAQ quantitation software (Quari) and measurement of and correction for isolation purity in iTRAQ
- Separation of peptides and phosphopeptide enrichment using IEF or OFF-Gel System
Instruments
Mass spectrometers
Thermo Orbitrap Velos Elite 1
Thermo Orbitrap Velos Elite 2
Thermo Orbitrap Excel with ETD
ABI 5800 MALDI-TOF/TOF
ABI 4000 Qtrap
2D-gel setup
Typhoon Scanner
Ettan Spot Handling Workstation
Ettan Spot Picker
Perkin Elmer Liquid Handling Station
Progenesis software programs
Supporting instruments
Three off-line Agilent HPLC for SCX/HILIC
OFFGEL Instrument for IEF separation of peptides and proteins
GelFree Instrument for in-solution separation of proteins
Data analysis
Workstations with Proteome Discoverer, Scaffold, Protein Pilot, Mascot Daemon, ProteoIQ
Core Staff
The Core staff members are experts in proteomics technologies. They will be happy to provide advice and training on equipment as necessary. If you would like to initiate a new project, please contact the Core Director Marjan Gucek, PhD via email.
Marjan Gucek
Core Director
Proteomics Core
P: +1 301 594 1060
F: +1 301 402 2113
gucekm@mail.nih.gov
Angel Aponte
Scientific Support Staff
Proteomics Core
P: +1 301 496 0004
apontea@mail.nih.gov
Yong Chen
Staff Scientist
Proteomics Core
P: +1 301 594 1187
yong.chen@nih.gov
Sajni Patel
Scientific Support Staff
Proteomics Core
P: +1 301 594 1272
F: +1 410 558 8054
patels2@mail.nih.gov
Stephen Swatkoski
Research Fellow
Proteomics Core
P: +1 301 594 1807
steve.swatkoski@nih.gov
Guanghui Wang
Staff Scientist
Proteomics Core
P: +1 301 402 2299
F: +1 301 402 2113
gw146x@nih.gov
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