Discrimination of Pathogenic vs. Nonpathogenic Francisella tularensis and Burkholderia pseudomallei Using Proteomics Mass Spectrometry
Abstract
The recent use of mass spectrometry (MS) analysis to characterize and identify biological agents using a proteomic approach has been demonstrated. The present study sought to determine whether proteomics MS could be used to distinguish between pathogenic and nonpathogenic strains of the same organism based on outer membrane proteins (OMPs). OMPs are often associated with virulence in gram-negative pathogens and could prove to be excellent model biomarkers for strain differentiation among bacteria. OMPs were isolated from pathogenic and nonpathogenic strains of Francisella tularensis and Burkholderia pseudomallei using ultracentrifugation and a sarkosyl extraction method. These methods are followed by protein digestion and analysis using liquid chromatography-MS/MS. The experimental MS/MS spectral data of bacterial peptides were searched using the SEQUEST algorithm against a constructed proteome database of microorganisms to determine the identity and number of unique peptides. Results showed that pathogenic strains could be distinguished from nonpathogenic strains for both organisms based on differences in OMP profiles. Virulent F. tularensis and B. pseudomallei expressed multiple OMPs that were not present in avirulent samples. In conclusion, proteomics MS proved useful in discriminating between pathogenic and non-pathogenic strains, and OMPs unique to pathogenic strains could prove to be useful biomarkers for strain differentiation.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 01, 2011
- Accession Number
- ADA540823
Entities
People
- Alan W. Zulich
- Charles H. Wick
- Mary M. Wade
- Rabih E. Jabbour
- Samir V. Deshpande
Organizations
- Edgewood Chemical Biological Center