The Experts below are selected from a list of 105 Experts worldwide ranked by ideXlab platform
Solomon H Snyder - One of the best experts on this subject based on the ideXlab platform.
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cloned and expressed macrophage nitric oxide synthase contrasts with the Brain Enzyme
Proceedings of the National Academy of Sciences of the United States of America, 1992Co-Authors: Charles J Lowenstein, Charles S Glatt, David S Bredt, Solomon H SnyderAbstract:Abstract Nitric oxide (NO) is a messenger molecule of macrophages, endothelial cells in blood vessels, and neurons. A neuronal form of NO synthase (NOS) has been previously cloned. We now report the molecular cloning of macrophage NOS. The macrophage Enzyme displays 50% sequence identity to the neuronal Enzyme. Like neuronal NOS, macrophage NOS has recognition sites for FAD, FMN, and NADPH and also has a consensus calmodulin binding site. Macrophage NOS mRNA is strikingly inducible; it is absent in quiescent macrophages or spleen but is prominent 2-6 hr after endotoxin treatment.
Shozo Yamamoto - One of the best experts on this subject based on the ideXlab platform.
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reversible hydrolysis and synthesis of anandamide demonstrated by recombinant rat fatty acid amide hydrolase
Biochemical and Biophysical Research Communications, 1997Co-Authors: Yuko Kurahashi, Natsuo Ueda, Hiroshi Suzuki, Mitsujiro Suzuki, Shozo YamamotoAbstract:Abstract Previously we suggested that one porcine Brain Enzyme (anandamide amidohydrolase) catalyzed both the hydrolysis of anandamide and its synthesis from arachidonic acid and ethanolamine (Ueda et al., J. Biol. Chem. 270, 23823–23827, 1995). In the present study we investigated the reversibility of the Enzyme reactions by the use of recombinant fatty-acid amide hydrolase of rat liver, which appears to be catalytically identical to porcine anandamide amidohydrolase. The particulate fraction of the COS-7 cells, in which the rat Enzyme was overexpressed, hydrolyzed anandamide with a specific activity of 132 nmol/min/mg protein at 37 °C, and the Km value for anandamide was 18 μM. The Enzyme also synthesized anandamide at a rate of 177 nmol/min/mg protein, and the Km values for arachidonic acid and ethanolamine as substrates were as high as 190 μM and 36 mM, respectively. The control cells transfected with the insert-free vector showed neither the hydrolase activity nor the synthase activity. Thus, the hydrolase and synthase are attributed to the same Enzyme protein coded by one gene. However, the Enzyme may act as a hydrolase rather than a synthase under physiological conditions judging from its high Km values for substrates in the synthase reactions. In addition, primary amides of fatty acids such as arachidonamide and oleamide and fatty acid ester like methyl arachidonate were hydrolyzed at considerable rates, and their reverse reactions occurred even if at lower rates.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Abstract Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 μmol/min/mg of protein at 37°C. As assayed with 14C-labeled substrates, the apparent K value for anandamide was 60 μM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 μmol/min/mg of protein at 37°C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 mumol/min/mg of protein at 37 degrees C. As assayed with 14C-labeled substrates, the apparent Km value for anandamide was 60 microM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 mumol/min/mg of protein at 37 degrees C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
Natsuo Ueda - One of the best experts on this subject based on the ideXlab platform.
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reversible hydrolysis and synthesis of anandamide demonstrated by recombinant rat fatty acid amide hydrolase
Biochemical and Biophysical Research Communications, 1997Co-Authors: Yuko Kurahashi, Natsuo Ueda, Hiroshi Suzuki, Mitsujiro Suzuki, Shozo YamamotoAbstract:Abstract Previously we suggested that one porcine Brain Enzyme (anandamide amidohydrolase) catalyzed both the hydrolysis of anandamide and its synthesis from arachidonic acid and ethanolamine (Ueda et al., J. Biol. Chem. 270, 23823–23827, 1995). In the present study we investigated the reversibility of the Enzyme reactions by the use of recombinant fatty-acid amide hydrolase of rat liver, which appears to be catalytically identical to porcine anandamide amidohydrolase. The particulate fraction of the COS-7 cells, in which the rat Enzyme was overexpressed, hydrolyzed anandamide with a specific activity of 132 nmol/min/mg protein at 37 °C, and the Km value for anandamide was 18 μM. The Enzyme also synthesized anandamide at a rate of 177 nmol/min/mg protein, and the Km values for arachidonic acid and ethanolamine as substrates were as high as 190 μM and 36 mM, respectively. The control cells transfected with the insert-free vector showed neither the hydrolase activity nor the synthase activity. Thus, the hydrolase and synthase are attributed to the same Enzyme protein coded by one gene. However, the Enzyme may act as a hydrolase rather than a synthase under physiological conditions judging from its high Km values for substrates in the synthase reactions. In addition, primary amides of fatty acids such as arachidonamide and oleamide and fatty acid ester like methyl arachidonate were hydrolyzed at considerable rates, and their reverse reactions occurred even if at lower rates.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Abstract Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 μmol/min/mg of protein at 37°C. As assayed with 14C-labeled substrates, the apparent K value for anandamide was 60 μM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 μmol/min/mg of protein at 37°C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 mumol/min/mg of protein at 37 degrees C. As assayed with 14C-labeled substrates, the apparent Km value for anandamide was 60 microM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 mumol/min/mg of protein at 37 degrees C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
Takashi Tokunaga - One of the best experts on this subject based on the ideXlab platform.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Abstract Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 μmol/min/mg of protein at 37°C. As assayed with 14C-labeled substrates, the apparent K value for anandamide was 60 μM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 μmol/min/mg of protein at 37°C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 mumol/min/mg of protein at 37 degrees C. As assayed with 14C-labeled substrates, the apparent Km value for anandamide was 60 microM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 mumol/min/mg of protein at 37 degrees C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
Yuko Kurahashi - One of the best experts on this subject based on the ideXlab platform.
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reversible hydrolysis and synthesis of anandamide demonstrated by recombinant rat fatty acid amide hydrolase
Biochemical and Biophysical Research Communications, 1997Co-Authors: Yuko Kurahashi, Natsuo Ueda, Hiroshi Suzuki, Mitsujiro Suzuki, Shozo YamamotoAbstract:Abstract Previously we suggested that one porcine Brain Enzyme (anandamide amidohydrolase) catalyzed both the hydrolysis of anandamide and its synthesis from arachidonic acid and ethanolamine (Ueda et al., J. Biol. Chem. 270, 23823–23827, 1995). In the present study we investigated the reversibility of the Enzyme reactions by the use of recombinant fatty-acid amide hydrolase of rat liver, which appears to be catalytically identical to porcine anandamide amidohydrolase. The particulate fraction of the COS-7 cells, in which the rat Enzyme was overexpressed, hydrolyzed anandamide with a specific activity of 132 nmol/min/mg protein at 37 °C, and the Km value for anandamide was 18 μM. The Enzyme also synthesized anandamide at a rate of 177 nmol/min/mg protein, and the Km values for arachidonic acid and ethanolamine as substrates were as high as 190 μM and 36 mM, respectively. The control cells transfected with the insert-free vector showed neither the hydrolase activity nor the synthase activity. Thus, the hydrolase and synthase are attributed to the same Enzyme protein coded by one gene. However, the Enzyme may act as a hydrolase rather than a synthase under physiological conditions judging from its high Km values for substrates in the synthase reactions. In addition, primary amides of fatty acids such as arachidonamide and oleamide and fatty acid ester like methyl arachidonate were hydrolyzed at considerable rates, and their reverse reactions occurred even if at lower rates.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Abstract Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 μmol/min/mg of protein at 37°C. As assayed with 14C-labeled substrates, the apparent K value for anandamide was 60 μM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 μmol/min/mg of protein at 37°C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.
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partial purification and characterization of the porcine Brain Enzyme hydrolyzing and synthesizing anandamide
Journal of Biological Chemistry, 1995Co-Authors: Natsuo Ueda, Yuko Kurahashi, Shozo Yamamoto, Takashi TokunagaAbstract:Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine Brain with 1% Triton X-100. The Enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 mumol/min/mg of protein at 37 degrees C. As assayed with 14C-labeled substrates, the apparent Km value for anandamide was 60 microM, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our Enzyme preparation. The purified Enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 mumol/min/mg of protein at 37 degrees C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single Enzyme protein.