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Approach to systematic analysis of serine/threonine phosphoproteome using Beta elimination and subsequent side effects: Intramolecular linkage and/or racemisation
Author(s) -
Tinette Sylvette,
Feyereisen René,
Robichon Alain
Publication year - 2006
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.21070
Subject(s) - dehydroalanine , serine , threonine , phosphorylation , biochemistry , chemistry , phosphopeptide , protein phosphorylation , trypsin , peptide , stereochemistry , protein kinase a , enzyme
Complete analysis of the phosphorylation of serine and threonine residues directly from biological extracts is still at an early stage and will remain a challenging goal for many years. Analysis of phosphorylated proteins and identification of the phosphorylated sites in a crude biological extract is a major topic in proteomics, since phosphorylation plays a dominant role in post‐translational protein modification. Beta elimination of the serine/threonine‐bound phosphate by alkali action generates (methyl)dehydroalanine. The reactivity of this group susceptible of nucleophilic attacks might be used as a tool for phosphoproteome analysis. Most of the known serine/threonine kinases recognize motifs in protein targets that are rich in lysine(s) and/or arginine(s). The (methyl)dehydroalanine resulting from beta elimination of the serine/threonine‐bound phosphate by alkali action is likely to react with the amino groups of these neighboring amino acids. Furthermore, the addition reaction of dehydroalanine‐peptides with a nucleophilic group more likely generates diastereoisomers derivatives. The internal cyclic bonds and/or the stereoisomer peptide derivatives thus generated confer resistance to trypsin cleavage and/or constitute stop signals for exopeptidases such as carboxypeptidase. This might form the basis of a method to facilitate the systematic identification of phosphorylated peptides. J. Cell. Biochem. 100: 875–882, 2007. © 2006 Wiley‐Liss, Inc.