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Enrichment of Bacterial Lipoproteins and Preparation of N-terminal Lipopeptides for Structural Determination by Mass Spectrometry
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Immunology and Infection
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JoVE Journal Immunology and Infection
Enrichment of Bacterial Lipoproteins and Preparation of N-terminal Lipopeptides for Structural Determination by Mass Spectrometry

Enrichment of Bacterial Lipoproteins and Preparation of N-terminal Lipopeptides for Structural Determination by Mass Spectrometry

DOI:
10.3791/56842-v

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10:59 min

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May 21, 2018

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,
1Department of Biochemistry and Molecular Biology,Pennsylvania State University

Chapters

  • 00:04Title
  • 00:47Cell Growth and Lysis
  • 02:23Enrichment of Lipoproteins by TX-114 Phase Partitioning
  • 05:05SDS-PAGE, Electroblotting, and Staining with Ponceau S
  • 06:30Tryptic Digestion, Lipopeptide Extraction from Nitrocellulose Membrane, Deposition onto MALDI Target, and Data Acquisition
  • 08:47Results: MALDI-TOF MS Analysis of N-terminal Lipopeptides
  • 10:17Conclusion

Summary

Automatic Translation

Automatic Translation

The enrichment of bacterial lipoproteins using a non-ionic surfactant phase partitioning method is described for direct use in TLR assays or other applications. Further steps are detailed to prepare N-terminal tryptic lipopeptides for structural characterization by mass spectrometry.

Tags

Keywords: Bacterial LipoproteinsN-terminal LipopeptidesStructural DeterminationMass SpectrometryMALDI-TOFToll-like ReceptorImmune ResponseTX-114 SurfactantPhase SeparationCell DisruptionZirconia Silica BeadsCentrifugation

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