Detection of S-nitrosylated protein by surface plasmon resonance

摘要

Abstract S-Nitrosylation has recently emerged as an important posttranslational modification of proteins and is becoming an intensive field of research in plants. Protein S-nitrosation, a reversible post-translation modification of cysteine, affects many cell signaling pathways and plays critical roles in redox-sensitive cell signaling. Changes in protein function effectively transmit biological signals and thus provide a framework for elucidating signaling networks. This paper presented a new, universal immunosensor for detection of S-nitrosylated proteins. Electrochemical impedance spectroscopy (EIS) and atomic force microscope (AFM) were used to estimate the formation of self-assembled film. This method was based on the specific binding characteristics of biotin–streptavidin, using Biotin-HPDP labeled protein sulfhydryl group as the substrate to detect proteins. The sensor was used to detect bovine serum albumin (BSA), nitrosylated {BSA} and denitrosylated BSA. The results showed that 90.61% of nitrosylated {BSA} were reduced, verifying that protein S-nitrosylation is a reversible and effective post-translation modification. This method was successfully applied to detect S-nitrosylated protein in Feicheng peach. The results showed good repeatability and precision. This method provided a molecular basis for further exploring the mechanism of S-nitrosylation of proteins in plants.