Table 1.
Overview of methodologies for detection of SNO-proteins.
Class 1: Detection of intact SNOa |
X-ray crystallography |
UV-vis spectroscopy |
NMR spectrocopy |
Mass spectrometry |
SNO-specific antibodies |
Class 2: Detection of SNO-derived nitrite and NOb |
Saville Assay |
DAF-2 Assay |
GC-MS |
Photolysis chemiluminscence |
Reductive chemiluminescence |
NO electrode |
Class 3: Labeling of SNO-derived Cys thiolc |
Biotin switch technique (BST) |
SNO-site identification (SNO-SID) |
S-nitrosothiol capture (SNOCAP) |
Resin-assisted capture of SNO-Proteins (SNO-RAC) |
Spin trapping after UV photolysis |
Organomercurial-binding |
Phosphine-based ligation |
These methods, except for SNO-based antibodies, are mostly suitable for the characterization of purified SNO-proteins and are generally low-sensitivity.
These methods are suitable for quantifying total levels of endogenous SNO-proteins but have limited utility for the analysis of specific SNO-proteins from complex mixtures.
These methods are useful for the enrichment, identification and relative quantification of SNO-proteins from complex mixtures and for the facile identification of SNO-sites.