Assessment of the nitrosylation process.

ABSTRACT

PURPOSE OF REVIEW To understand the principles and limits of the methodologies used for the measurement of S-nitrosylated proteins. RECENT FINDINGS: Among methods for studying protein S-nitrosylation, chemoluminescence and biotin switch assay have rapidly gained popularity. However, recent findings have attempted to highlight potential pitfalls for these methods. Many assays for biological S-nitrosylated proteins are used near the limit of detection and pretreatment of the biological samples can modify the S-NO bond. These results suggest that additional controls are essential in order to identify S-nitrosylated proteins and results should be quantitatively validated using more than one methodology. SUMMARY: Protein S-nitrosylation is emerging as a key mechanism by which nitric oxide regulates cell signalling. This review focuses on existing methodologies for the measurement of S-nitrosylated proteins in biological matrices and the potential pitfalls of each method.