Cysteine tagging for MS‐based proteomics

Abstract

Amino acid-tagging strategies are widespread in proteomics. Because of the central role of mass spectrometry (MS) as a detection technique in protein sciences, the term "mass tagging" was coined to describe the attachment of a label, which serves MS analysis and/or adds analytical value to the measurements. These so-called mass tags can be used for separation, enrichment, detection, and quantitation of peptides and proteins. In this context, cysteine is a frequent target for modifications because the thiol function can react specifically by nucleophilic substitution or addition. Furthermore, cysteines present natural modifications of biological importance and a low occurrence in the proteome that justify the development of strategies to specifically target them in peptides or proteins. In the present review, the mass-tagging methods directed to cysteine residues are comprehensively discussed, and the advantages and drawbacks of these strategies are addressed. Some concrete applications are given to underline the relevance of cysteine-tagging techniques for MS-based proteomics.

The SNO/SOH TMT strategy for combinatorial analysis of reversible cysteine oxidations

“…For effective analysis of SNO and SOH selective labelling and enrichment are required [13]. A number of methods for analysis of cysteine oxidation exist, each with certain advantages and limitations, as reviewed in [2].…”

A Novel Strategy for Global Analysis of the Dynamic Thiol Redox Proteome

“…Numerous techniques have been developed because of the increasing interest and evidences of redox-regulated signaling networks (8,9). For a long time, the determination of the redox-regulated functional role of specific Cys sites has been performed using site-directed mutagenesis (10).…”

Implementing visible 473 nm photodissociation in a Q-Exactive mass spectrometer: towards specific detection of cysteine-containing peptides

“…22 Specific photodissociation was also obtained for Tyr/His-containing peptides after derivatization with diazonium chromophore groups. 23 Detection specificity is also critical when tackling protein quantification in whole proteome by targeted mass spectrometry in selected reaction monitoring mode (SRM 25 Thus, photo-SRM showed similar or even improved detection performance compared to conventional CID-SRM for the analysis of a whole plasma hydrolysate.…”