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Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
Published on: February 20, 2013
Proteolytic 18O-labeling strategies for quantitative proteomics.
Masaru Miyagi1, K C Sekhar Rao
1Case Center for Proteomics, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106, USA. masaru.miyagi@case.edu
Mass Spectrometry Reviews
|November 7, 2006
Summary
Proteolytic 18O-labeling is a quantitative proteomic technique that uses a protease and 18O-water to label peptides for protein quantification. This method enables comparative proteomics, analysis of post-translational modifications, and protein interaction studies.
Area of Science:
- Biochemistry
- Proteomics
- Analytical Chemistry
Background:
- Quantitative proteomics is essential for understanding biological systems.
- Various techniques exist for protein quantification, each with limitations.
- Proteolytic 18O-labeling offers a specific approach to relative protein quantification.
Purpose of the Study:
- To review the proteolytic 18O-labeling technique in detail.
- To discuss its history, advantages, disadvantages, and applications.
- To cover advancements, enzymatic considerations, and data analysis.
Main Methods:
- Utilizes a protease and H(2)18O for peptide labeling.
- Employs chromatographic and mass spectrometric analysis for identification and quantification.
- Determines relative protein expression ratios between samples.
Main Results:
- The technique allows for relative quantification of proteins.
- It is applicable to comparative proteomics and post-translational modification studies.
- Recent advancements address variable 18O incorporation issues.
Conclusions:
- Proteolytic 18O-labeling is a valuable tool for quantitative proteomics.
- The review provides a comprehensive overview of the technique and its biological applications.
- Further advancements enhance its reliability and scope.
