Volume 22, Issue 96 (12-2012)
J Mazandaran Univ Med Sci 2012, 22(96): 59-69 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Kadkhodaei Elyaderani M, Malek Askar A M, Rostami M, Aberomand M, Khirollah A. Kinetic Activity of Isolated Nitric-oxide Synthase Enzyme from Sheep Kidney. J Mazandaran Univ Med Sci 2012; 22 (96) :59-69
URL: http://jmums.mazums.ac.ir/article-1-1702-en.html
URL: http://jmums.mazums.ac.ir/article-1-1702-en.html
Manizheh Kadkhodaei Elyaderani 
, Ali Mohammad Malek Askar , Morad Rostami , Mohammad Aberomand , Alireza Khirollah
, Ali Mohammad Malek Askar , Morad Rostami , Mohammad Aberomand , Alireza Khirollah
Abstract: (11186 Views)
Background and purpose: Nitric oxide (NO) is produced by nitric oxide synthase (NOS) from L-arginine and has an important role in a variety of physiological and pathological processes in biological systems. In this study, kinetic activities of isolated NOS were evaluated from sheep kidney.
Materials and methods: In this reasearch homogenization, ammonium sulfate precipitation and column chromatography on DEAE-32 Cellulose and 2', 5'-ADP-agarose of 500 grams of sheep kidney were used to purify isolated nitric oxide. In all steps of purification process the amount of protein and its activity was assayed using Bradford and Griess reactions.
Results: The molecular weight of sheep kidney on SDS-PAGE electrophoresis was 54 KD. Specific activity was 0.6 units/mg protein and recovery of purification was 0.9% (relative purity was
20 times more). Optimum temperature and pH were 30˚C and 7.4 and incubation of purified enzyme at thermal interval of 10 to 30˚C for 15 minute showed a stable enzyme activity. At 75 μM concentration of L-arginine substrate, the highest level of NOS activity was seen. The Vmax and Km of enzyme were
250 nmol/min/mg protein and 5.32 M, respectively.
Conclusion: NOS enzyme from sheep kidney was purified with relative purity of 20 times and their optimum temperature, pH, suitable substrate concentration, Vmax and Km were 30˚C, 7.4, 75 μM, 250 nmol/min/mg proteins and 5.32 µmol, respectively.
Type of Study: Research(Original) |
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
Articles Copyright © The Author(s). Owned by Mazandaran University of Medical Sciences.
Contact Us
Address: Journal Office, Building No. 2, Mazandaran University of Medical Sciences, Moallem Square, Sari.Tel: +98 (011)34484874 Postal code: 48175-866 E-Mail: jmums@mazums.ac.ir Telefax: +98 (011)33262679
