This paper reports a systemic mass spectrometry (MS) investigation of a novel strategy for labeling biological thiols, involving the cleavage of the Se-N bond by thiol to form a new Se-S bond. Our data show that the reaction is highly selective, rapid, reversible, and efficient. Among 20 amino acids, only cysteine is reactive toward Se-N containing reagents and the reaction occurs in seconds. With the addition of dithiothreitol, peptides derivatized by selenium reagents can be recovered. The high reaction selectivity and reversibility provide potential in both selective identification and isolation of thiols from mixtures. Also, with dependence on the selenium reagent used, derivatized peptide ions exhibit tunable dissociation behaviors (either facile cleavage or preservation of the formed Se-S bond upon collision-induced dissociation), a feature that is useful in proteomics studies. Equally importantly, the thiol derivatization yield is striking, as reflected by 100% conversion of protein β-lactoglobulin A using ebselen within 30 s. In addition, preliminary applications such as rapid screening of thiol peptides from mixtures and identification of the number of protein free and bound thiols have been demonstrated. The unique selenium chemistry uncovered in this study would be valuable in the MS analysis of thiols and disulfide bonds of proteins/peptides.
All Science Journal Classification (ASJC) codes
- Analytical Chemistry