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David R. Goodlett, Ph.D., Director, Proteomics
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Overview
The Proteomics component of the Genetics Core assists CHDD Research Affiliates through an arrangement with the Department of Medicinal Chemistry. This collaboration puts Center investigators in contact with Goodlett's proteomics facility, to plan proteomics experiments and have their samples analyzed.
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Location
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Faculty & Staff
David R. Goodlet, Ph.D.
Director, Proteomics
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Byron Gallis, Ph.D.
Research Supervisor
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To Use Our Services
Investigators who are interested in using the proteomics facility in their projects should:
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Services & Equipment
The overall objective of the Proteomics component of the Genetics Core is to provide state-of-the-art capabilities for identifying and characterizing proteins via mass spectrometry. Services include:
- peptide sequencing,
- shotgun proteomics,
- comparative proteomics using the isotope-coded affinity tag (ICAT) labeling method,
- identification of post-translational modifications.
- qualitative or descriptive analysis of proteomes,
- relative quantification of changes in proteomes induced by various environmental or genetic perturbations,
- peptide sequence analysis, and
- identification of post-translational modifications to proteins.
These services fit into two types of experimental protocols: (1) hypothesis testing/targeted analysis by mass spectrometry (MS) and (2) molecular modeling, hypothesis-generating, or discovery-based analysis.
More specifically, the Proteomics Facility provides CHDD Research Affiliates access to descriptive proteomic analysis via MS as well as tandem MS (MS/MS). These methods include excision of identified 1-D or 2-D protein stained “spots” as well as shotgun proteomic methods, in which no or minimal separation is done at the level of proteins prior to MS analysis. The purpose is to identify proteins by sequence match in a database or by de novo derivation using MS methods.
The Proteomics Facility also provides support to determine changes in proteome expression resulting from select biochemical/environmental perturbations as well as genetic differences. This is carried out using the quantitative shotgun proteomic approach with ICAT and multidimensional liquid chromatography (MDLC) separation of peptides.
Support is also provided for the determination of protein post-translational modifications (PTMs). These experiments use both (1) low-energy collision-induced dissociation (CID) on an electrospray ionization quadrupole time-of-flight (ESI-QTOF) MS and (2) electron capture dissociation (ECD) on Fourier transform–ion cyclotron resonance–MS (FT-ICR-MS). The purpose is to sequence peptides suspected to be modified by a PTM and to locate/identify the PTM(s).
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Fourier transform–ion cyclotron
resonance–mass spectrometer (4.7 T). |
Staff provide consultation and training of investigators and the operation of complex equipment such as mass spectrometers. Investigators are responsible for preparation of their own samples and the costs associated with preparation. However, sample preparation will be collaborative and done under the supervision of Dr. Goodlett’s staff.
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