Selective liberation of NO from S-nitrosocysteine with potassium thiocyanate, as monitored by an amperometric sensor.
S-Nitrosocysteine (CysNO) releases either NO (in the presence of divalent cations) or NO+ (in the presence of chelating agents). NO+ is then transferred to peptides or protein SH groups to form high-mass nitrosothiols. The aim of this work was the development of a specific reaction between thiocyanate (SCN-) and CysNO. This reaction selectively liberates NO from CysNO in the presence of high-mass nitrosothiols. Free NO is measured with an amperometric sensor. We examine with this system the transnitrosylation reaction between CysNO and BSA at low molecular ratios and could assay nitrites, SNO-BSA, and CysNO in the incubation mixture without any preliminary purification steps.
Alberto
Palmerini Carlo
PC
Dipartimento di Scienze Biochimiche e Biotecnologie Molecolari, Via del Giochetto, 06127 Perugia, Italy. crlpal@unipg.it
Marmottini
Fabio
F
Arienti
Giuseppe
G
Palombari
Roberto
R
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004396:Coloring Agents / Q000737:chemistry; D003545:Cysteine / Q000031:analogs & derivatives* / Q000737:chemistry*; D003553:Cystine / Q000737:chemistry; D004594:Electrophysiology; D009569:Nitric Oxide; D020030:Nitric Oxide Donors / Q000737:chemistry; D009573:Nitrites / Q000737:chemistry; D026403:S-Nitrosothiols / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013861:Thiocyanates / Q000494:pharmacology*; D013997:Time Factors
D004396:Coloring Agents; D020030:Nitric Oxide Donors; D009573:Nitrites; D026403:S-Nitrosothiols; D013861:Thiocyanates; D012710:Serum Albumin, Bovine; D009569:Nitric Oxide; D003553:Cystine; C021315:S-nitrosocysteine; D003545:Cysteine; C009941:potassium thiocyanate