Assessment of the nitrosylation process.
OBJECTIVE
To understand the principles and limits of the methodologies used for the measurement of S-nitrosylated proteins.


RESULTS
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.


CONCLUSIONS
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.
Borderie
Didier
D
Laboratoire de biochimie A, Hôpital Cochin APHP, 27 rue du faubourg Saint Jacques, France. didier.borderie@cch.aphp.fr
Ekindjian
Ohvanesse G
OG
D001710:Biotin / Q000378:metabolism; D015153:Blotting, Western; D003124:Colorimetry; D003545:Cysteine / Q000378:metabolism; D008163:Luminescent Measurements; D013058:Mass Spectrometry; D009569:Nitric Oxide / Q000378:metabolism*; D015538:Nitrosation; D011506:Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013439:Sulfhydryl Reagents
D011506:Proteins; D013439:Sulfhydryl Reagents; D009569:Nitric Oxide; D001710:Biotin; D003545:Cysteine