A novel approach to identify proteins modified by nitric oxide: the HIS-TAG switch method.
S-nitrosylation is emerging as an important signaling mechanism that regulates a broad range of cellular functions. The recognition of Cysteine residues that undergo S-nitrosylation is crucial to elucidate how this modification modulates protein activity. We report here a novel strategy, defined His-tag switch, which allows the purification and identification of S-nitrosylated proteins and the unambiguous localization of the modified cysteine residues by mass spectrometry analysis.
Camerini
Serena
S
San Raffaele Scientific Institute, 20132 Milan, Italy. serena.camerini@iss.it
Polci
Maria L
ML
Restuccia
Umberto
U
Usuelli
Vera
V
Malgaroli
Antonio
A
Bachi
Angela
A
D000595:Amino Acid Sequence; D000818:Animals; D002531:Cerebellum / Q000378:metabolism*; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000737:chemistry; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism*; D009603:Nitroso Compounds / Q000737:chemistry*; D010047:Ovalbumin / Q000032:analysis / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D026422:S-Nitrosoglutathione / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D009419:Nerve Tissue Proteins; D009603:Nitroso Compounds; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D010047:Ovalbumin