Speaker: Dr. Melinda McFarland, FDA/Center forFood Safety and Applied Nutrition
Topic: Top-Down Mass Spectrometry for the Rapid Identification of Strain Specific Bacterial Proteins
Date: Monday, February 13, 2012
Time: 7:30 pm
Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)
Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm. Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.
Abstract: Faster and more effective methods for bacterial assessment are becoming increasingly relevant. Intact protein expression profiling by LCMS is a powerful tool for bacterial strain differentiation. However, transfer to field usable assays or monitoring of drift in these species requires identification of marker proteins. The availability of predicted proteins from a rapidly expanding database of bacterial genomes affords the possibility of developing tools in which partial protein sequence information obtained from top-down mass spectrometry can rapidly identify strain specific proteins. Combined with the associated molecular weight information, the approach can identify variations in protein sequence that are related to bacterial strain differences. In addition, comparative proteogenomics of closely related bacteria by top-down MSMS allows us to go beyond identification of possible cross-serovar orthologs. Access to intact protein masses, HPLC retention times, and site specific MSMS data provides insight into specific mutations, PTMs, start site errors and signal peptide cleavages that are often poorly annotated in sequence databases. However, intact protein analysis does present its own set of limitations. Data will also be presented illustrating why the current state of intact protein analysis generally yields significantly fewer protein masses than are actually present in the bacteria.