一氧化氮分子在細胞間訊息傳遞可藉由與半胱胺酸上的硫基形成可逆共價鍵結，稱為硫基亞硝基化/去亞硝基化。目前已有許多文獻指出蛋白質的活性和功能可經由此種調節進而影響多種生理機制。然而，S-NO鍵的不穩定增加了研究上的挑戰，使得目前對於亞硝基化的條件和反應機制仍不明確。現今發展的許多偵測RSNO的方法，可直接或間接的提供部分RSNO訊息，其中應用最廣泛的為類生物素轉換技術，此方法可經由一系列覆蓋策略將RSNO從細胞樣品中純化與鑑定。
2009年，Wang 和Xian等人發表了一種可將一級RSNO轉換成相對穩定的雙硫鍵產物反應機制，稱為二次接合反應，此反應可在溫和的條件下進行，且獲得良好的產率。基於此研究，我們將硫酯基探針與磁性奈米粒子結合，應用於純化和鑑定RSNOs。將亞硝基化標準胜肽PTP1B與BSA複雜樣品混合後，磁性探針可成功的純化出PTP1B，證實了此想法的可行性，我們相信此探針在研究亞硝基化蛋白上具有極大的潛力。

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.

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