Abstract
Nitric oxide (NO) influenced on cellular signaling in large part by means of S-nitrosylation/denitrosylation, the NO group covalently attach to the thiol side chain of cysteine resulting in the formation of S-nitrosothiols (RSNOs). RSNOs regulated proteins activity and function in a wide range of cellular pathways and physiological processes. However, the condition and mechanism of RSNOs formation remains uncertain, due to the lability of the S-NO bond caused the challenging for research. To date, numerous methods for assay RSNOs directly or indirectly have been developed. Among methods for studying RSNOs, biotin switch technology (BST) was used most commonly which can be used to detect and isolate SNO proteins from cell extracts in series of capping steps. In 2009, Wang and Ming et al. reported a bis-ligation mechanism of RSNOs which can converts unstable primary RSNOs to stable disulfide-iminophosphorane products in high yields under mild conditions. Based on this study, we conjugated the thioester phosphine ligand to the magnetic nanoparticle(MNP) for identification and purification of RSNOs. To verify the feasibility of the idea, the S-nitrosylated PTP1B peptide mixed in tryptic BSA mixture (50 ug) was successfully captured by the magnet probe and identified by MALDI-TOF. We believe this strategy wiil become a potential tool used in investigate S-nitrosylation proteins.