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The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes
Published on: April 10, 2015
High sequence-coverage detection of proteolytic peptides using a bis(terpyridine)ruthenium(II) complex
Akihiro Ito1, Taka-Aki Okamura, Ken Masui
1Department of Macromolecular Science, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.
The Analyst
|June 8, 2007
Summary
This study introduces Ru-CO labeling for mass spectrometry, enhancing peptide detection by overcoming common interferences. This novel method improves sensitivity and accuracy in protein analysis, even in complex mixtures.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Peptide labeling is crucial for mass spectrometry (MS) based protein identification.
- Traditional methods can be affected by protonation or sodiated adduction, complicating analysis.
- Developing sensitive and reliable labeling techniques is essential for advancing proteomics.
Purpose of the Study:
- To evaluate a bis(terpyridine)ruthenium(II) complex for peptide labeling (Ru-CO labeling) in MS.
- To assess the impact of Ru-CO labeling on peptide detection sensitivity and accuracy.
- To demonstrate the applicability of Ru-CO labeling for complex protein mixture analysis.
Main Methods:
- Utilized a bis(terpyridine)ruthenium(II) complex for Ru-CO labeling of peptides.
- Employed matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for analysis.
- Applied matrix-assisted laser desorption/ionization post-source-decay (MALDI-PSD) for sequence determination.
- Tested the method on insulin, myoglobin, and a mixture of myoglobin, lysozyme, and ubiquitin.
Main Results:
- Ru-CO labeling produced high-intensity peaks in MS, overcoming protonation/sodiation interferences.
- Simultaneous detection of labeled insulin A- and B-chains with comparable abundance was achieved.
- MALDI-PSD enabled easy amino acid sequence determination of labeled peptide fragments.
- Sensitivity was independent of peptide length or sequence.
- Tryptic myoglobin fragments showed near-equal abundance, aiding identification above 670.70 Da.
- Achieved 96% sequence coverage for myoglobin via peptide mass fingerprinting (PMF).
- Successfully characterized a protein mixture (>90% sequence coverage) simultaneously.
Conclusions:
- Ru-CO labeling significantly enhances peptide detection in mass spectrometry.
- The method offers high sensitivity, accuracy, and applicability to complex biological samples.
- This technique provides a powerful tool for proteomic analysis and protein characterization.
