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Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis
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JoVE Journal Chemistry
Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis

Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis

DOI:
10.3791/62577-v

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09:26 min

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May 23, 2021

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,
1Produce Safety & Microbiology, Western Regional Research Center,Agricultural Research Service, U.S. Department of Agriculture

Chapters

  • 00:04Introduction
  • 00:40Mass Spectrometry
  • 03:23Operating Protein Biomarker Seeker Software
  • 06:57Results: Analysis of Bacterial Proteins Using Mass Spectrometry
  • 08:57Conclusion

Summary

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Here we present a protocol for the rapid identification of proteins produced by genomically sequenced pathogenic bacteria using MALDI-TOF-TOF tandem mass spectrometry and top-down proteomic analysis with software developed in-house. Metastable protein ions fragment because of the aspartic acid effect and this specificity is exploited for protein identification.

Tags

Keywords: Escherichia ColiAntibacterial Immunity ProteinsMALDI-TOF-TOF-MS/MSTop-down ProteomicsMass SpectrometryBacterial Protein IdentificationAntibiotic InductionSample PreparationHPLC-grade WaterSinapinic AcidMS Linear Mode AcquisitionMS/MS Reflectron Mode AcquisitionPrecursor MassIsolation WidthCIDMetastable Suppressor

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