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Updated: Jun 11, 2025
09:43
Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
Published on: February 20, 2013
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Enzymatic Synthesis of Isotopically Labeled Hydrogen Peroxide for Mass Spectrometry-Based Applications
Margaret Hoare1, Ruiyue Tan1, Isabella Militi1
1Department of Biology, University of Rochester, Rochester, New York 14627, United States.
Journal of the American Society for Mass Spectrometry
|October 4, 2024
Summary
Researchers developed an affordable in-house method to synthesize isotopically labeled hydrogen peroxide (H218O2). This advancement facilitates accurate measurement of methionine oxidation in vivo using mass spectrometry.
Area of Science:
- Biochemistry
- Proteomics
- Chemical Biology
Background:
- Methionine oxidation plays a role in protein misfolding and enzyme regulation.
- Measuring methionine oxidation in vivo is challenging due to artifactual oxidation during sample preparation and mass spectrometry.
- Isotopically labeled hydrogen peroxide (H218O2) can accurately measure in vivo methionine oxidation but is costly and difficult to source.
Purpose of the Study:
- To develop an accessible in-house synthesis method for H218O2.
- To validate the use of enzymatically synthesized H218O2 for accurate methionine oxidation measurements.
Main Methods:
- Enzymatic synthesis of H218O2 using glucose oxidase and 18O2.
- Assessment of synthesized H218O2 concentration and purity.
- Application of synthesized H218O2 in mass spectrometry workflows for methionine oxidation analysis.
Main Results:
- A reliable method for in-house H218O2 synthesis was established.
- The synthesized H218O2 was characterized for concentration and purity.
- The utility of the synthesized H218O2 in mass spectrometry for accurate methionine oxidation quantification was demonstrated.
Conclusions:
- In-house synthesis of H218O2 provides a cost-effective alternative for researchers.
- This method enables accurate measurement of in vivo methionine oxidation levels.
- The developed protocol supports broader research into the biological roles of methionine oxidation.
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