Synthesis and analytical investigation of C‐terminally modified peptide aldehydes and ketone: application to oxime ligation |
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Authors: | Corinne Buré Philippe Marceau Hervé Meudal Agnès F Delmas |
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Institution: | 1. Centre de Biophysique Moléculaire CNRS UPR4301, affiliated to the University of Orléans and INSERM, , 45071 Orléans cedex 2, France;2. Present address: Chimie et Biologie des Membranes et des Nanoobjets (CBMN) – UMR 5248 Centre de Génomique Fonctionnelle BP 68, Université Bordeaux 2 Victor Segalen, , 33076 Bordeaux Cedex, France |
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Abstract: | C‐terminally modified peptides aldehyde (glycinal and alpha‐oxo aldehyde peptides) and ketone (pyruvic acid‐containing peptide) were synthesised to get new insights into the mechanism of acido‐catalysed oxime ligation. Their tetrahedral hydrated forms were investigated in solution and in the gas phase, using NMR and in‐source collision‐induced dissociation mass spectrometry, respectively, and the kinetics of the oximation reactions followed using analytical HPLC. The results obtained confirmed that the first step of the oximation reaction was the limiting step for the pyruvic acid‐containing peptides because of the steric effect and of the carbon angular strain of the ketone. The second step is the determining step for the aldehyde peptides because the basicity of the oxygen of the hydroxyl function of the tetrahedral form is greater for glycinal than for alpha‐oxo aldehyde. These data strongly suggest that the hydrated form of the aldehyde partner has to be considered when oxime reactions are performed in aqueous buffer. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd. |
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Keywords: | peptide aldehyde peptide ketone ESI/mass spectrometry NMR oxime ligation HPLC kinetic |
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