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
peer-reviewed · Analytical Chemistry · 2000. Martin Münchbach et al. We describe a method for comparative quantitation and de novo peptide sequencing of proteins separated either…
| 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 | 7.10 |
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 (28)
- Bromine isotopic signature facilitates de novo sequencing of peptides in free‐radical‐initiated peptide sequencing (FRIPS) mass spectrometry (2015) crossref
- Lessons in de novo peptide sequencing by tandem mass spectrometry (2015) both
- Neutron-encoded Signatures Enable Product Ion Annotation From Tandem Mass Spectra (2013) both
- Differential 14N/15N-Labeling of Peptides Using N-Terminal Charge Derivatization with a High-Proton Affinity for Straightforward de novo Peptide Sequencing (2013) crossref
- Stable Isotope N-Phosphorylation Labeling for Peptide de Novo Sequencing and Protein Quantification Based on Organic Phosphorus Chemistry (2012) crossref
- 193 nm Ultraviolet Photodissociation of Imidazolinylated Lys-N Peptides for De Novo Sequencing (2012) crossref
- De novo analysis of protein N-terminal sequence utilizing MALDI signal enhancing derivatization with Br signature (2012) crossref
- Improving peptide fragmentation by N‐terminal derivatization with high proton affinity (2011) crossref
- Dimethyl isotope labeling assisted de novo peptide sequencing (2010) crossref
- De novo sequencing of peptides by MS/MS (2010) crossref
- Peptide de Novo Sequencing Using 157 nm Photodissociation in a Tandem Time-of-Flight Mass Spectrometer (2010) crossref
- N-terminal tagging strategy forDe Novosequencing of short peptides by ESI-MS/MS and MALDI-MS/MS (2010) both
- Acetylating Tryptic Peptides Enhances b Ion Intensity in MALDI TOF/TOF: Implications in Peptide Sequencing and Identification of Proteins in an Antarctic Bacterium Pseudomonas Syringae (2010) both
- De novo sequence analysis and intact mass measurements for characterization of phycocyanin subunit isoforms from the blue‐green alga Aphanizomenon flos‐aquae (2009) crossref
- Rapid and Reliable Peptide de Novo Sequencing Facilitated by Microfluidic Chip-Based Edman Degradation (2008) crossref
- N-Terminal amino acid side-chain cleavage of chemically modified peptides in the gas phase: A mass spectrometry technique for N-terminus identification (2006) 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
- Peptide de Novo Sequencing Facilitated by a Dual-Labeling Strategy (2005) crossref
- Enhancing TOF/TOF-based de Novo Sequencing Capability for High Throughput Protein Identification with Amino Acid-Coded Mass Tagging (2005) crossref
- Isotope‐coded N‐terminal sulfonation of peptides allows quantitative proteomic analysis with increased de novo peptide sequencing capability (2004) crossref
- Two-Dimensional Mass Spectra Generated from the Analysis of 15N-Labeled and Unlabeled Peptides for Efficient Protein Identification and de novo Peptide Sequencing (2004) crossref
- A case study of de novo sequence analysis of N-sulfonated peptides by MALDI TOF/TOF mass spectrometry (2004) 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
- Precise peptide sequencing and protein quantification in the human proteome through in vivo lysine-specific mass tagging (2003) crossref
- Use of Deuterium-Labeled Lysine for Efficient Protein Identification and Peptide de Novo Sequencing (2002) crossref
- De novo peptide sequencing and quantitative profiling of complex protein mixtures using mass-coded abundance tagging (2002) crossref