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Constitutive nitric oxide synthase in Saccharomyces cerevisiae
R N Kanadia1, W N Kuo, M Mcnabb
1Division of Science and Mathematics, Bethune-Cookman College, Daytona Beach, Florida 32114, USA.
Biochemistry and Molecular Biology International
|October 8, 1998
Summary
This study detected constitutive nitric oxide synthase (cNOS) in yeast, demonstrating its presence in unicellular eukaryotes. The enzyme
Area of Science:
- Biochemistry
- Molecular Biology
- Yeast Genetics
Background:
- Nitric oxide synthase (NOS) enzymes are crucial in various biological processes.
- The presence and function of NOS in unicellular eukaryotes like yeast have been largely unexplored.
Purpose of the Study:
- To investigate the occurrence and characteristics of constitutive nitric oxide synthase (cNOS) in Saccharomyces cerevisiae (yeast).
- To determine if yeast cNOS exhibits enzymatic activity and how it is regulated.
Main Methods:
- Immunoaffinity chromatography and Western blotting were used to detect and identify yeast cNOS.
- Enzyme activity was assessed by monitoring citrulline formation.
- The effects of calmodulin, arginine, and L-NAME on enzyme activity were evaluated.
- The impact of actinomycin-D on nitrate and nitrite levels was measured.
Main Results:
- A 60 kDa immunoreactive band corresponding to cNOS was identified in yeast extracts.
- Yeast cNOS activity was confirmed, showing activation by calmodulin and arginine, and inhibition by L-NAME.
- Actinomycin-D treatment reduced nitrate and nitrite levels, indicating NO production regulation.
Conclusions:
- Constitutive nitric oxide synthase (cNOS) is present in unicellular eukaryotes such as yeast.
- Yeast cNOS activity is regulated by specific factors, similar to mammalian NOS.
- This finding expands our understanding of NOS distribution and function across different life forms.