Partial Purification and Characterization of the Porcine Brain Enzyme Hydrolyzing and Synthesizing Anandamide (*)
Partial Purification and Characterization of the Porcine Brain Enzyme Hydrolyzing and Synthesizing Anandamide (*)
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猪脑酶水解和合成 Anandamide 的部分纯化和表征 (*)
DOI：
10.1074/jbc.270.40.23823
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
T. Tokunaga
中科院分区：
文献类型：
--
作者：
N. Ueda;Y. Kurahashi;Shozo Yamamoto;T. Tokunaga
Anandamide（花生四烯乙醇酰胺）被认为是大麻素受体的内源性激动剂。用1%Triton X-100从猪脑微粒体部分中溶解一种水解花生四烯酸的酰胺水解酶。通过Phenyl-5 PW疏水色谱法将酶部分纯化至在37°C下约0.37 μmol/min/mg蛋白质的比活性。用14 C标记的底物测定，花生四烯酸的表观K值为60 μM，花生四烯酸的活性高于亚油酸、油酸和棕榈酸的乙醇酰胺。在我们的酶制剂中未检测到神经酰胺酶和蛋白酶活性。纯化的酶还在高浓度乙醇胺存在下从游离花生四烯酸合成花生四烯酸，在37°C下比活性约为0.16 μmol/min/mg蛋白质。的基础上的共色谱，pH值的依赖性，热灭活，和抑制剂，如花生四烯三氟甲基酮，对氯汞苯甲酸，二异丙基氟磷酸盐，和苯甲基磺酰氟的影响，有人建议，花生四烯酸酰胺水解酶和合成酶的活动是由于一个单一的酶蛋白。
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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影响因子：
5.8
作者：
S. Childers;T. Sexton;Mary Beth Roy
通讯作者：
S. Childers;T. Sexton;Mary Beth Roy
Anandamide 是一种内源性大麻素，作为 N18 神经母细胞瘤细胞的部分激动剂抑制钙电流。
影响因子：
3.6
作者：
Mackie,K;Devane,WA;Hille,B
通讯作者：
Hille,B
DOI：
10.1073/pnas.90.16.7656
发表时间：
1993-08-15
影响因子：
11.1
作者：
FELDER, CC;BRILEY, EM;DEVANE, WA
通讯作者：
DEVANE, WA
anandamide（一种假定的内源性大麻素）在小鼠中的药理活性。
DOI：
--
发表时间：
1994
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Smith,PB;Compton,DR;Welch,SP;Razdan,RK;Mechoulam,R;Martin,BR
通讯作者：
Martin,BR
影响因子：
56.9
作者：
DEVANE, WA;HANUS, L;MECHOULAM, R
通讯作者：
MECHOULAM, R