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Chapters
- 00:05Title
- 01:43PALeO-TimeCourse: Protease-catalyzed Labeling of Proteolytic Cleavages
- 02:43PALeO-TimeCourse: Postdigestion Labeling of Proteolytic Termini
- 03:48ALeO-TimeCourse: Acid-catalyzed Labeling of Carboxyl Groups
- 04:25MALDI-TOF/TOF MS/MS Data Acquisition and Analysis
- 06:35Preparation of Spectral Time and 18O-incorporation Plots
- 07:06Representative Results
- 08:58Conclusion
Stable isotope labeling workflows employing 18O-enriched water (LeO-workflows) are versatile tools for quantitative and qualitative proteomics studies. In protease-assisted (PALeO) workflows, 18O-atoms are introduced by proteolytic cleavage and carboxyl oxygen exchange reactions mediated by proteases. In the acid-catalyzed (ALeO) workflow, 18O-atoms are introduced by carboxyl oxygen exchange at low pH.
Tags
Protease-catalyzed LabelingAcid-catalyzed Labeling18O-enriched WaterStable IsotopesBiological Mass SpectrometryCatalytic MechanismsProteolytic EnzymesMass Spectrometry-based ProteomicsProtein Expression Levels18O-labelingPeptide Bond CleavageCarboxyl Oxygen Exchange ReactionPALeO AdaptationEnzymatic 18O-labelingDistinctive Isotope SignaturesHigh-resolution Matrix-assisted Laser Desorption Ionization Time-of-flight Tandem Mass Spectrometry (MALDI-TOF/TOF MS/MS)Isotope Envelopes
