The rate of protein S-nitrosylation, a reversible process by which S-nitroso thiol (RS-NO) compounds exchange the NO+ moiety with protein SH groups, is essentially governed by two factors, the pK alpha and the accessibility of the protein sulfhydryl. A useful method of following transnitrosation kinetics of various protein and nonprotein SH compounds with GS-NO is described. When the reaction is carried out in the presence of 1-chloro-2,4-dinitrobenzene and glutathione transferases, the rate of RS-NO formation (RSH + GS-NO-->RS-NO + GSH) can be monitored by spectrophotometry at 340 nm in terms of the enzymatic conversion of GSH to a GS conjugate. Unlike methods based on NO release from the S-NO bond, this procedure is rapid and accurate and requires relatively small amounts of thiols. The second order rate constants of S-nitrosylation of human and rat oxy- and deoxyhemoglobin of BSA and other thiols were calculated by this method which confirmed previous results reported in the literature

Rossi, R., Lusini, L., Giannerini, F., Giustarini, D., Lungarella, G., DI SIMPLICIO, P. (1997). A method to study kinetics of transnitrosation with nitrosoglutathione: reactions with hemoglobin and other thiols. ANALYTICAL BIOCHEMISTRY, 254(2), 215-220 [10.1006/abio.1997.2424].

A method to study kinetics of transnitrosation with nitrosoglutathione: reactions with hemoglobin and other thiols

ROSSI, R.;GIUSTARINI, D.;LUNGARELLA, G.;DI SIMPLICIO, P.
1997

Abstract

The rate of protein S-nitrosylation, a reversible process by which S-nitroso thiol (RS-NO) compounds exchange the NO+ moiety with protein SH groups, is essentially governed by two factors, the pK alpha and the accessibility of the protein sulfhydryl. A useful method of following transnitrosation kinetics of various protein and nonprotein SH compounds with GS-NO is described. When the reaction is carried out in the presence of 1-chloro-2,4-dinitrobenzene and glutathione transferases, the rate of RS-NO formation (RSH + GS-NO-->RS-NO + GSH) can be monitored by spectrophotometry at 340 nm in terms of the enzymatic conversion of GSH to a GS conjugate. Unlike methods based on NO release from the S-NO bond, this procedure is rapid and accurate and requires relatively small amounts of thiols. The second order rate constants of S-nitrosylation of human and rat oxy- and deoxyhemoglobin of BSA and other thiols were calculated by this method which confirmed previous results reported in the literature
Rossi, R., Lusini, L., Giannerini, F., Giustarini, D., Lungarella, G., DI SIMPLICIO, P. (1997). A method to study kinetics of transnitrosation with nitrosoglutathione: reactions with hemoglobin and other thiols. ANALYTICAL BIOCHEMISTRY, 254(2), 215-220 [10.1006/abio.1997.2424].
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11365/21412
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