Quantitation and Facilitated de Novo Sequencing of Proteins by Isotopic N-Terminal Labeling of Peptides with a Fragmentation-Directing Moiety
peer-reviewed · Analytical Chemistry · 2000
| Date | 2000-09-01 |
| Type | peer-reviewed |
| Venue | Analytical Chemistry |
| Publisher | American Chemical Society (ACS) |
| Contribution | adjacent |
| DOI | 10.1021/ac000265w |
| Citations (OpenAlex) | 280 |
| Venue 2-year citedness | 6.29 |
Abstract
We describe a method for comparative quantitation and de novo peptide sequencing of proteins separated either by standard chromatographic methods or by one- and two-dimensional polyacrylamide gel electrophoresis. The approach is based on the use of an isotopically labeled reagent to quantitate (by mass spectrometry) the ratio of peptides from digests of a protein being expressed under different conditions. The method allows quantitation of the changes occurring in spots or bands that contain more than one protein and has a greater dynamic range than most staining methods. Since the reagent carries a fixed positive charge under acidic conditions and labels only the N-terminal of peptides, the interpretation of tandem mass spectra to obtain sequence information is greatly simplified. The sequences can easily be extracted for homology searches instead of using indirect mass spectral-based searches and are independent of posttranslational modifications.
Methods and tools
- Isotopic N-terminal labeling de novo sequencing: Isotopic N-terminal labeling with a fragmentation-directing moiety for protein quantitation and facilitated de novo sequencing.
Cites (1)
Cited by (8)
- Lessons in de novo peptide sequencing by tandem mass spectrometry (2015) both
- Dimethyl isotope labeling assisted de novo peptide sequencing (2010) crossref
- De novo sequencing of peptides by MS/MS (2010) crossref
- De Novo Sequencing of Neuropeptides Using Reductive Isotopic Methylation and Investigation of ESI QTOF MS/MS Fragmentation Pattern of Neuropeptides with N-Terminal Dimethylation (2005) crossref
- De Novo Peptide Sequencing Based on a Divide-and-Conquer Algorithm and Peptide Tandem Spectrum Simulation (2004) crossref
- “De novo” peptide sequencing by MALDI-quadrupole-ion trap mass spectrometry: A preliminary study (2003) crossref
- PEAKS: powerful software for peptide de novo sequencing by tandem mass spectrometry (2003) crossref
- De novo peptide sequencing and quantitative profiling of complex protein mixtures using mass-coded abundance tagging (2002) crossref