Effect of Genetic Database Comprehensiveness on Fractional Proteomics of Escherichia coli O157:H7
Abstract
This report is required for the U.S. Army Edgewood Chemical Biological Center (ECBC) In-House Laboratory Independent Research project "Membrane Vesicles and Extracellular Proteins in Antibiotic Resistance and Virulence." The binding of extracellular proteins [membrane vesicle (MV) or secreted] to antibiotics could contribute toward antibiotic resistance mechanisms. We are characterizing the extracellular, fimbriae, and whole cell proteins produced by the pathogenic Gram-negative bacterium Escherichia coli (E. coli) O157:H7 in terms of proteomics and antibiotic-binding using mass spectrometry. Here we report a study on the effect of the comprehensive nature of the database used for proteomics on the analysis of various protein fractions. In this case, analysis using a more restricted database (chosen based on sample knowledge) resulted in identification of a higher percentage of proteins than a more comprehensive database. For E. coli O157:H7, we identified proteins that were specific to certain cellular fractions. Of these, four penicillin binding proteins were identified solely in the secretome fraction. Although the three penicillin binding proteins (PBPs) whose functions have been determined are antibiotic targets for penicillin and they therefore do not play an antibiotic resistance role, the identification of PBPs solely in the secretome does agree with our hypothesis that antibiotic proteins would be observed in the extracellular fraction.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2014
- Accession Number
- ADA591867
Entities
People
- Mary M. Wade
- Patrick E. Mccubbin
- Rabih E. Jabbour
- Samir V. Deshpande
- Vicky L.H. Bevilacqua
Organizations
- Edgewood Chemical Biological Center