NMR detection and study of hydrolysis of HNO-derived sulfinamides.
Nitroxyl (HNO), a potential heart failure therapeutic, is known to post-translationally modify cysteine residues. Among reactive nitrogen oxide species, the modification of cysteine residues to sulfinamides [RS(O)NH2] is unique to HNO. We have applied (15)N-edited (1)H NMR techniques to detect the HNO-induced thiol to sulfinamide modification in several small organic molecules, peptides, and the cysteine protease, papain. Relevant reactions of sulfinamides involve reduction to free thiols in the presence of excess thiol and hydrolysis to form sulfinic acids [RS(O)OH]. We have investigated sulfinamide hydrolysis at physiological pH and temperature. Studies with papain and a related model peptide containing the active site thiol suggest that sulfinamide hydrolysis can be enhanced in a protein environment. These findings are also supported by modeling studies. In addition, analysis of peptide sulfinamides at various pH values suggests that hydrolysis becomes more facile under acidic conditions.
Department of Chemistry, Johns Hopkins University , Baltimore, Maryland 21218, United States.
Gizem
G
Keceli
Cathy D
CD
Moore
Jason W
JW
Labonte
John P
JP
Toscano
D000577:Amides / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D009589:Nitrogen Oxides / Q000737:chemistry*; D010084:Oxidation-Reduction; D010206:Papain / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D013441:Sulfinic Acids / Q000737:chemistry*
D000577:Amides; D009589:Nitrogen Oxides; D010446:Peptide Fragments; D013441:Sulfinic Acids; D010206:Papain; C039900:nitroxyl; D003545:Cysteine