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Design and synthesis of a solid-phase fluorescent mass tag
Yang Shi1, Rong Xiang, Csaba Horváth
1Department of Chemical Engineering, Yale University, New Haven, CT, USA.
Journal of Separation Science
|October 18, 2005
Summary
We developed a new fluorescent mass tag (FMT) for peptide labeling. This cost-effective method uses stable isotopes and fluorescence detection for improved protein analysis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Proteomics
Background:
- Solid-phase synthesis is crucial for efficient peptide modification.
- Stable isotope labeling is a powerful tool in quantitative proteomics.
- Fluorescence detection offers high sensitivity in biomolecular analysis.
Purpose of the Study:
- To design and synthesize a novel solid-phase fluorescent mass tag (FMT).
- To enable cost-effective and efficient labeling of cysteine-containing peptides.
- To facilitate sensitive detection of labeled peptides using fluorescence and mass spectrometry.
Main Methods:
- Solid-phase synthesis utilizing Fmoc chemistry.
- Uniform labeling of lysine with stable isotopes for mass difference.
- Coupling of a fluorescent molecule to the isotope tag.
- Incorporation of an iodoacetyl reactive group for cysteine specificity.
- Analysis using MALDI-TOF MS and HPLC.
Main Results:
- Successful synthesis of the FMT reagent.
- Demonstrated 8 Da mass difference between heavy and light tags.
- Effective labeling of standard cysteine peptides.
- Successful labeling of cysteine-containing peptides from a BSA tryptic digest.
- Online fluorescence detection of tagged peptides.
Conclusions:
- The developed FMT is a versatile and efficient tool for peptide labeling.
- The combination of stable isotope labeling, fluorescence, and mass spectrometry enhances peptide analysis.
- This method offers a cost-effective approach for quantitative proteomics applications.