Ferrous cytochrome c-nitric oxide oxidation for quantification of protein S-nitrosylation probed by resonance Raman spectroscopy

Protein S-nitrosylation is an important post-translational modification, which has important effect on protein functions and is relevant to various diseases. For quantification of the S-nitrosated proteins, it is still challenging to establish a detection method in combination of fast and one-step determination, highly sensitivity and accuracy. Here, we develop a label-free, direct detection method for S-nitrosated proteins based on resonance Raman scattering of ferrous cytochrome c. The laser source of the Raman spectrometer is utilized both for inducing NO leakage and for Raman scattering excitation. The heme ligation alteration and oxidation enable fast quantification of the released NO by the resonance Raman spectroscopy of the cytochrome c. All the results shown in this study demonstrate the great potential of the proposed method for fast, sensitive and reliable assessment of total S-nitrosated proteins in the clinical applications.