Identification of Protein Interactions via Mass Spectroscopy
Produce Safety and Microbiology Research
2011 Annual Report
1a.Objectives (from AD-416)
The ultimate goal of this project is to identify proteins of interest in certain diseases chosen for study by Prosetta and ARS, and any locations of protein-protein and protein-ligand binding relevant to the mechanisms of those diseases, for use inter alia in understanding a mechanism of action (MOA) between the protein and a particular ligand to develop new diagnostics, therapeutics, and prophylactics.
1b.Approach (from AD-416)
Prosetta will provide ARS with putative protein targets that Prosetta will purify using a variety of affinity techniques; the proteins may include various modifications and cross-links. Final purification will be done by SDS-PAGE gels that will be stained with coomassie blue. The proteins of interest will be cut from the gels are then processed to yield a set of peptides, which be analyzed by mass spectrometry to identify the sequences and structural features of the proteins. Further analysis will be performed to reveal the location of any modifications or crosslinks.
In the second year of this collaboration we have received samples from Prosetta and identified proteins by mass spectrometry. This data and its analysis have been transferred to Prosetta and are considered proprietary.