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Lysine‐directed staining of proteins for MS ‐based analyses
Author(s) -
Yasui Matthew T.,
MataGómez Marco A.,
Winkler Robert
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200438
Subject(s) - lysine , chemistry , cysteine , peptide , covalent bond , proteome , combinatorial chemistry , biochemistry , computational biology , amino acid , biology , organic chemistry , enzyme
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 U niblue A drastically speeds up proteomic workflows. The present study introduces dabsyl chloride as another truly MS ‐compatible protein stain. Remarkably, although U niblue 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 AL i PHAT strategy for increasing the sensitivity of peptide identifications.