Protein thiol denoted as ‘RSH’. GSNO reacting with the thiol can produce either S-nitrosylation (RSNO) or S-glutathionylation (RSSG), depending on the conditions and the particular thiol involved. Treatment with NO• donor, SNAP, produces only S-nitrosylation. Successive treatments with dithiodipyridine (DTDP) and GSH produces glutathionylation in a two step reaction: DTDP oxidizes the thiol, leaving a pyridine group (Pyr) linked to the protein by a disulphide bond (RSSPyr), and subsequent treatment with GSH produces RSSG by a thiol-disulphide exchange reaction. DTT reduces both RSNO and RSSG back to RSH (not shown).