Inhibitory effect of asymmetric dimethylarginine and NG-Monomethyl-L-arginine methyl ester on nitric oxide synthase activity

Autor: Kadkhodaei Elyaderani M, Malek Askar AM, Rostami M, Aberomand M, Kheirollah AR
Jazyk: perština
Rok vydání: 2013
Předmět:
Zdroj: مجله دانشگاه علوم پزشکی گرگان, Vol 15, Iss 3, Pp 84-92 (2013)
Druh dokumentu: article
ISSN: 1562-4765
2008-4080
Popis: Background and Objective: Nitric oxide synthase (NOS) paly a role in nitric oxide (NO) generation. Despite the beneficial effects of NO on different body systems its overproduction of produce reactive nitrogen species (RNS) and nitrosilation of proteins. This study was done to evaluate the effect of asymmetric dimethylarginine (ADMA) and NG-Monomethyl-L-arginine methyl ester (L-NMMA) on inhibition of nitric oxide synthase activity. Materials and Methods: In this laboratory study, Nitric oxide synthase was extracted from 500 grams of sheep kidney by homogenization, ammonium sulphate precipitation and column chromatography on DEAE-32 Cellulose and 2', 5'-ADP-agarose. During purification, protein content was measured according to the Bradford and enzyme activity was assayed using the Griess reactions the inhibitory effects of 25 μΜ concentrations of ADMA and L-NMMA on purified enzyme were determined. Results: Specific activity and yield of NOS were 0.6 units/mg protein and 0.9%, respectively. Molecular weight of purified enzyme was 54 KD with SDS-PAGE. ADMA and L-NMMA in 25 μΜ concentrations reduced enzyme activity by 76 and 61.2%, respectively. Km values for NOS in absence and in presence of ADMA and L-NMMA were 5.32 μM, 31.25 μM (P
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