Related Experiment Video
Updated: Jun 20, 2026
10:37
Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
Measuring proteome dynamics in vivo: as easy as adding water?
Nadia Rachdaoui1, Leanne Austin, Eric Kramer
1Department Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.
Molecular & Cellular Proteomics : MCP
|September 3, 2009
Summary
This study introduces labeled water as a novel method for quantifying proteome dynamics, offering a more practical and reliable approach for measuring protein synthesis in vivo compared to traditional labeled amino acids.
Area of Science:
- Proteomics and Systems Biology
- Metabolic Research
- Biochemical Assays
Background:
- Proteomics typically provides static gene expression data, limiting the study of dynamic proteome changes.
- Challenges in studying proteome dynamics include in vivo isotope administration, low protein labeling detection, and interpreting precursor/product labeling.
- Current methods often rely on labeled amino acids, which have limitations in long-term studies and precursor labeling interpretation.
Purpose of the Study:
- To demonstrate the potential of quantifying proteome dynamics using stable isotopes and mass spectrometry.
- To evaluate labeled water (2H2O vs H218O) as an alternative to labeled amino acids for measuring in vivo protein synthesis.
- To highlight the advantages of labeled water for both short-term and long-term proteomic studies.
Main Methods:
- Coupling stable isotope administration with mass spectrometric assays.
- Comparing the use of labeled water (2H2O and H218O) versus labeled amino acids for measuring albumin biosynthesis in vivo.
- Utilizing precursor/product labeling ratios for rate calculations in protein synthesis studies.
Main Results:
- Labeled water enables the in vivo generation of labeled amino acids, overcoming limitations of continuous labeled amino acid administration in long-term studies.
- Labeled water simplifies precursor labeling measurement, crucial for accurate protein synthesis rate calculations in short-term studies.
- Demonstrated successful application of labeled water for studying albumin synthesis in mice under both steady-state and non-steady-state metabolic conditions.
Conclusions:
- Labeled water offers a practical and reliable method for quantifying proteome dynamics and in vivo protein synthesis.
- This approach overcomes key challenges associated with traditional labeled amino acid methods.
- Labeled water is broadly applicable for large-scale investigations of proteome dynamics, providing functional insights into gene expression.
