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Enzymes for anandamide biosynthesis and metabolism
N Ueda1, Y Kurahashi, K Yamamoto
1Department of Biochemistry, Tokushima University, School of Medicine, Japan.
Journal of Lipid Mediators and Cell Signalling
|September 1, 1996
Summary
Researchers isolated and purified anandamide amidohydrolase, an enzyme that breaks down anandamide. This enzyme also synthesizes anandamide, suggesting a single protein performs both functions.
Area of Science:
- Biochemistry
- Neuroscience
- Enzymology
Background:
- Anandamide is an endogenous ligand that interacts with cannabinoid receptors.
- Understanding the enzymes involved in anandamide metabolism is crucial for neuroscience research.
Purpose of the Study:
- To isolate and purify the enzyme responsible for hydrolyzing anandamide (anandamide amidohydrolase).
- To investigate the enzymatic properties and potential dual functions of this enzyme.
Main Methods:
- Enzyme isolation from porcine brain microsomal fraction.
- Solubilization using Triton X-100.
- Partial purification via hydrophobic chromatography.
- Enzyme activity assays including K(m) determination for anandamide.
- Inhibitor experiments to assess enzyme function.
Main Results:
- Anandamide amidohydrolase activity was localized in the brain microsomal fraction.
- The enzyme was partially purified, exhibiting a specific activity of 0.3 mumol/min per mg protein.
- The apparent K(m) for anandamide was approximately 60 microM.
- The enzyme demonstrated lower activity with other fatty acid ethanolamides.
- The same enzyme preparation exhibited anandamide synthase activity, converting arachidonic acid to anandamide.
Conclusions:
- A single enzyme protein appears to possess both anandamide amidohydrolase and anandamide synthase activities.
- This finding provides new insights into the metabolic regulation of anandamide in the brain.