Cysteine-reactive covalent capture tags for enrichment of cysteine-containing peptides

Article Article scientifique Article Cysteine-reactive covalent capture tags for enrichment of cysteine-containing peptides eng 2009 author Giron Priscille 632225 author Dayon Loic 652150 author Mihala Nikolett 624857 author Sanchez Jean-Charles 0000-0002-8733-5932 24399 author Rose Keith 21876 3377-3386 10.1002/rcm.4261 19813279 0951-4198 Considering the tremendous complexity and the wide dynamic range of protein samples from biological origin and their proteolytic peptide mixtures, proteomics largely requires simplification strategies. One common approach to reduce sample complexity is to target a particular amino acid in proteins or peptides, such as cysteine (Cys), with chemical tags in order to reduce the analysis to a subset of the whole proteome. The present work describes the synthesis and the use of two new cysteinyl tags, so-called cysteine-reactive covalent capture tags (C3T), for the isolation of Cys-containing peptides. These bifunctional molecules were specifically designed to react with cysteines through iodoacetyl and acryloyl moieties and permit efficient selection of the tagged peptides. To do so, a thioproline was chosen as the isolating group to form, after a deprotection/activation step, a thiazolidine with an aldehyde resin by the covalent capture (CC) method. The applicability of the enrichment strategy was demonstrated on small synthetic peptides as well as on peptides derived from digested proteins. Mass spectrometric (MS) analysis and tandem mass spectrometric (MS/MS) sequencing confirmed the efficient and straightforward selection of the cysteine-containing peptides. The combination of C3T and CC methods provides an effective alternative to reduce sample complexity and access low abundance proteins. Animals Cross-Linking Reagents/*chemistry Cysteine/analysis/*chemistry Horses Humans Mass Spectrometry/*methods Myoglobin/analysis/chemistry Peptide Fragments/analysis/*chemistry Proteins/analysis/*chemistry Serum Albumin/analysis/chemistry Thiazolidines/chemistry Dewey 576 136 Département de biologie structurale et bioinformatique 21E false 369 Biomarqueurs 635 RCM. Rapid communications in mass spectrometry 23 21 9051e9d7-b085-4bea-af78-ae982ef4b786 Article (Published version) Restricted file1.pdf 400215 application/pdf restrictedAccess Faculté de médecine / Section de médecine fondamentale / Département de biologie structurale et bioinformatique

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Cysteine-reactive covalent capture tags for enrichment of cysteine-containing peptides	RCM. Rapid communications in mass spectrometry			2009	637	3

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Mihala, Nikolett