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Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry
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Biology
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JoVE Journal Biology
Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry

Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry

DOI:
10.3791/3766-v

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12:07 min

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March 24, 2012

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, , ,
1Plant Molecular and Cellular Biology Program,University of Florida , 2Department of Biology,University of Florida , 3Interdisciplinary Center for Biotechnology Research,University of Florida , 4Genetics Institute,University of Florida

Chapters

  • 00:05Title
  • 02:07Inoculation of Tomato
  • 03:13Protein Extraction
  • 04:21Sample Preparation and Peptide Labeling with cysTMT
  • 06:07Removal of Non-reacted Tag Sample Fractionation
  • 07:44Sample Enrichment of cysTMT-labeled Peptides
  • 08:43Mass Spectrometry Analysis
  • 10:02CysTMT Results: Profiling Thiol Redox Proteome of Pseudomonas syringae in Tomato
  • 11:11Conclusion

Summary

Automatic Translation

Automatic Translation

Reactive oxygen species level is elevated when cells encounter stress conditions. Here we show the example of 3'-3' diaminobenzidine staining as well as cysTMT labeling and mass spectrometry to profile the redox proteome in Pseudomonas syringae treated tomato leaves.

Tags

Thiol Redox ProteomeIsotope Tagging Mass SpectrometryPseudomonas Syringae Pv. TomatoBacterial Speck DiseaseSolanum LycopersicumBrassica SpeciesArabidopsis ThalianaReactive Oxygen Species (ROS)Hydrogen PeroxideDAB StainingRedox Status Of ProteinsCysteine OxidationTandem Mass Tag (TMT) ReagentsCysteine-reactive TMT (cysTMT) Tags

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