Differential stable isotope labeling of peptides for quantitation and de novo sequence derivation
peer-reviewed · Rapid Communications in Mass Spectrometry · 2001
| Date | 2001-07-30 |
| Type | peer-reviewed |
| Venue | Rapid Communications in Mass Spectrometry |
| Publisher | Wiley |
| Contribution | adjacent |
| DOI | 10.1002/rcm.362 |
| Citations (OpenAlex) | 281 |
| Venue 2-year citedness | 1.80 |
Abstract
We have demonstrated the use of per-methyl esterification of peptides for relative quantification of proteins between two mixtures of proteins and automated de novo sequence derivation on the same dataset. Protein mixtures for comparison were digested to peptides and resultant peptides methylated using either d0- or d3-methanol. Methyl esterification of peptides converted carboxylic acids, such as are present on the side chains of aspartic and glutamic acid as well as the carboxyl terminus, to their corresponding methyl esters. The separate d0- and d3-methylated peptide mixtures were combined and the mixture subjected to microcapillary high performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). Parent proteins of methylated peptides were identified by correlative database searching of peptide tandem mass spectra. Ratios of proteins in the two original mixtures could be calculated by normalization of the area under the curve for identical charge states of d0- to d3-methylated peptides. An algorithm was developed that derived, without intervention, peptide sequence de novo by comparison of tandem mass spectra of d0- and d3-peptide methyl esters.
Methods and tools
- Differential methyl esterification labeling: Labels peptide carboxyl groups with d0- or d3-methanol so that mixed samples give quantitation and isotope-paired fragment ions that support automated de novo sequencing.
Cites (3)
Cited by (13)
- Enhancing De Novo Protein Sequencing through the C-Terminal Labeling Strategy: Resolving Isobaric Ambiguities by Electron-Transfer/Higher Energy Collision Dissociation (EThcD) (2024) crossref
- Neutron-encoded Signatures Enable Product Ion Annotation From Tandem Mass Spectra (2013) both
- C-terminal de novo sequencing of peptides using oxazolone-based derivatization with bromine signature (2011) crossref
- Rapid Validation of Mascot Search Results via Stable Isotope Labeling, Pair Picking, and Deconvolution of Fragmentation Patterns (2009) both
- 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
- Improved peptide sequencing using isotope information inherent in tandem mass spectra (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