Electrophoresis 2013-02-01

Lysine-directed staining of proteins for MS-based analyses.

Matthew T Yasui, Marco A Mata-Gómez, Robert Winkler

Index: Electrophoresis 34(3) , 401-4, (2013)

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Abstract

Visualization of proteins and MS-based analyses are elemental tasks in modern biochemistry. Nevertheless, reports about covalent protein dyes and their suitability for subsequent MS experiments remain scarce. In a recent work, we demonstrated that covalent prestaining of proteins with Uniblue A drastically speeds up proteomic workflows. The present study introduces dabsyl chloride as another truly MS-compatible protein stain. Remarkably, although Uniblue A and dabsyl chloride employ different nucleophilic reaction mechanisms, both are highly specific for lysine residues. The predictable peptide modifications allow easy integration into state-of-the-art bioinformatic workflows. Further, lysine-directed derivatizations with hydrophobic reagents such as dabsyl chloride complement the cysteine-directed ALiPHAT strategy for increasing the sensitivity of peptide identifications.© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.