Chemical methods to detect S-nitrosation

Abstract

Nitric oxide (NO) is a cell-signaling molecule involved in a number of physiological and pathophysiological processes. Modification of cysteine residues by NO (or NO metabolites), that is S-nitrosation, changes the function of a broad spectrum of proteins. This reaction represents an important post-translational modification that transduces NO-dependent signals. However, the detection and quantification of S-nitrosation in biological samples remain a challenge mainly because of the lability of S-nitrosation products: S-nitrosothiols (SNO). In this review we summarize recent developments of the methods to detect S-nitrosation. Our focus is on the methods which can be used to directly conjugate the site(s) of S-nitrosation.

Figure 1

Common methods for SNO detection

Figure 2

Biotin switch based methods for SNO detection. (a) Original biotin switch. (b) Resin assisted capture. (c) Fluorescence switch. (d) Microarray based assay. R shown here indicates thiol blocking groups.

Figure 3

Phosphine based methods for SNO detection. (a) Reductive ligation. (b) Bis-ligation. (c) One-step disulfide formation. (d) Reductive elimination. (e) S-alkylphosphonium formation. R shown here indicates different S-nitrosothiol substrates, mainly cysteine derivatives. R′ indicates aryl or alkyl substituent groups.