ZFIN ID: ZDB-PUB-081114-18
A versatile peptide pI calculator for phosphorylated and N-terminal acetylated peptides experimentally tested using peptide isoelectric focusing
Gauci, S., van Breukelen, B., Lemeer, S.M., Krijgsveld, J., and Heck, A.J.
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
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.
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