PUBLICATION
A versatile peptide pI calculator for phosphorylated and N-terminal acetylated peptides experimentally tested using peptide isoelectric focusing
- Authors
- Gauci, S., van Breukelen, B., Lemeer, S.M., Krijgsveld, J., and Heck, A.J.
- ID
- ZDB-PUB-081114-18
- Date
- 2008
- Source
- Proteomics 8(23-24): 4898-4906 (Journal)
- Registered Authors
- Lemeer, Simone
- Keywords
- N-terminal acetylation, Peptide IEF, Phosphorylation, pI calculator, Titanium oxide
- MeSH Terms
-
- Acetylation
- Animals
- Cell Extracts
- Isoelectric Focusing/methods*
- Isoelectric Point
- Phosphopeptides/chemistry*
- Phosphorylation
- Reagent Strips
- Zebrafish/embryology
- Zebrafish/metabolism
- PubMed
- 19003858 Full text @ Proteomics
Citation
Gauci, S., van Breukelen, B., Lemeer, S.M., Krijgsveld, J., and Heck, A.J. (2008) A versatile peptide pI calculator for phosphorylated and N-terminal acetylated peptides experimentally tested using peptide isoelectric focusing. Proteomics. 8(23-24):4898-4906.
Abstract
We experimentally demonstrate the use of an in-house developed pI calculator which takes into account peptide PTM such as phosphorylation and N-terminal acetylation. The pI calculator was utilized for a large set of peptides derived from a complex zebrafish lysate fractionated using peptide IEF, whereby a good correlation between the calculated (theoretical) pI and the experimental pI could be established. This pI calculator permits the implementation of optimal pK values depending on the experimental conditions and a reliable calculation of peptide pI which can be utilized as a filtering technique in validating peptide identifications. Our data reveal that the shift due to a phosphorylation or N-terminal acetylation is highly dependent on the presence of acidic or basic residues in the peptide. Furthermore, using this pI calculator, we revealed previously unknown position-specific pKs of asparagine and carbamidomethylated cysteine depending on their location in the peptide. Collectively, this peptide pI calculator is a welcome addition to the versatility and robustness of IEF for the separation and confident identification of (post-translationally modified) peptides.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping