De novo sequence analysis of N‐terminal sulfonated peptides after in‐gel guanidination
peer-reviewed · PROTEOMICS · 2005
| Date | 2005-06-01 |
| Type | peer-reviewed |
| Venue | PROTEOMICS |
| Publisher | Wiley |
| Contribution | algorithm |
| DOI | 10.1002/pmic.200501311 |
| Citations (OpenAlex) | 28 |
Abstract
Here we report a novel approach in which gel-separated proteins are guanidinated in-gel prior to enzymatic cleavage. In contrast to previously described techniques, this procedure allows the extracted tryptic peptides to be N-terminal sulfonated without any further sample purification. The derivatized peptides were subsequently fragmented using a matrix-assisted laser desorption/ionization time of flight/time of flight instrument. The approach facilitates the de novo sequence analysis and allows obtaining longer stretches of amino acid sequence information. We demonstrate that the obtained information can be used to identify proteins using a sequence similarity search algorithm. The technique was compared to the standard peptide mass fingerprint approach, applied either in-gel or in solution, using a number of sodium dodecyl sulfate-polyacrylamide gel electrophoresis separated model proteins. Finally, the new protocol was applied on a proteomic study of two-dimensional PAGE separated proteins from Shewanella oneidensis. More than 50 proteins from this organism were identified using sub-picomol quantities of protein, and peptide sequences of up to 20 amino acid residues in length have been determined.
Methods and tools
- N-sulfonated peptide MALDI TOF/TOF de novo sequencing: N-sulfonation before MALDI TOF/TOF directs fragmentation into a single dominant series, so a peptide mass fingerprint that does not identify a protein on its own can be followed by a readable de novo spectrum.
Cites (5)
- A case study of de novo sequence analysis of N-sulfonated peptides by MALDI TOF/TOF mass spectrometry (2004) crossref
- De novo sequencing of peptides using MALDI/TOF-TOF (2002) crossref
- De novo peptide sequencing and quantitative profiling of complex protein mixtures using mass-coded abundance tagging (2002) crossref
- Derivatization procedures to facilitate de novo sequencing of lysine-terminated tryptic peptides using postsource decay matrix-assisted laser desorption/ionization mass spectrometry (2000) crossref
- A method for high-sensitivity peptide sequencing using postsource decay matrix-assisted laser desorption ionization mass spectrometry (1999) crossref
Cited by (9)
- De novo analysis of protein N-terminal sequence utilizing MALDI signal enhancing derivatization with Br signature (2012) crossref
- The use of 2D-electrophoresis and de novo sequencing to characterize inter- and intra-cultivar protein polymorphisms in an allopolyploid crop (2011) crossref
- Biomolecular characterization of allergenic proteins in snow crab (Chionoecetes opilio) and de novo sequencing of the second allergen arginine kinase using tandem mass spectrometry (2011) crossref
- Characterization and de novo sequencing of snow crab tropomyosin enzymatic peptides by both electrospary ionization and matrix‐assisted laser desorption ionization QqToF tandem mass spectrometry (2010) crossref
- Simplifying Fragmentation Patterns of Multiply Charged Peptides by N-Terminal Derivatization and Electron Transfer Collision Activated Dissociation (2009) crossref
- De novo sequence analysis and intact mass measurements for characterization of phycocyanin subunit isoforms from the blue‐green alga Aphanizomenon flos‐aquae (2009) crossref
- Lys Tag: an easy and robust chemical modification for improved de novo sequencing with a matrix‐assisted laser desorption/ionization tandem time‐of‐flight mass spectrometer (2008) crossref
- Functional Proteome Analysis of the Banana Plant (Musa spp.) Using de Novo Sequence Analysis of Derivatized Peptides (2007) crossref
- MALDI‐TOF/TOF de novo sequence analysis of 2‐D PAGE‐separated proteins from Halorhodospira halophila, a bacterium with unsequenced genome (2006) crossref