Reversible Hydrolysis and Synthesis of Anandamide Demonstrated by Recombinant Rat Fatty-Acid Amide Hydrolase

Kurahashi, Yuko; Ueda, Natsuo; Suzuki, Hiroshi; Suzuki, Mitsujiro; Yamamoto, Shozo

doi:10.1006/bbrc.1997.7180

Cited by 93 publications

(60 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…More interestingly, the enzyme was significantly inhibited by AEA, the preferential substrate of FAAH [10], and ATFMK or MAFP, two inhibitors of this enzyme [9,11]. FAAH was previously shown to be expressed in both cell types under study, to display a pHdependency profile identical with that shown in Figure 1(C) for [$H]2-AG hydrolysis [8,17] and to catalyse the hydrolysis of arachidonoyl methyl ester [12]. Oleamide and palmitoylethanol-2-Arachidonoylglycerol inactivation in whole cells and cell-free preparations (Figures 2A and 2B) to an extent very similar to that previously observed for their inhibition of FAAH respectively in N18TG2 and RBL-2H3 cells [8,17].…”

Section: Resultsmentioning

confidence: 86%

“…This enzyme is inhibited by classical serine-and cysteine-protease inhibitors, such as N-ethylmaleimide (NEM) and p-hydroxymercuribenzoate (pHMB) [7][8][9][10], as well as by AA derivatives such as arachidonoyltrifluoromethyl ketone (ATFMK) and arachidonoylmethyl fluorophosphonate (MAFP) [9,11]. When overexpressed in COS-7 cells, FAAH was shown to catalyse also the hydrolysis of the sleep-inducing factor cis-9-octadecenoamide (oleamide) [10] and of arachidonoyl methyl ester [12]. Only a few metabolic studies have been performed to date on 2-AG [13][14][15], some of which have shown potential connections between its formation and AEA production [13].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The novel endogenous cannabinoid 2-arachidonoylglycerol is inactivated by neuronal- and basophil-like cells: connections with anandamide

Marzo

Bisogno

Sugiura

et al. 1998

Biochemical Journal

191

View full text Add to dashboard Cite

The novel endogenous cannabinoid 2-arachidonoylglycerol (2-AG) was rapidly inactivated by intact rat basophilic leukaemia (RBL-2H3) and mouse neuroblastoma (N18TG2) cells through diffusion\hydrolysis\reacylation processes. The hydrolysis of 2-AG was inhibited by typical esterase inhibitors and by more specific blockers of ' fatty acid amide hydrolase ' (FAAH), the enzyme catalysing the hydrolysis of the other ' endocannabinoid ', anandamide (AEA). No evidence for a facilitated-diffusion process was found. A 2-AG-hydrolysing activity was detected in homogenates from both cell lines, with the highest levels in membrane fractions. It exhibited an optimal pH at 10, and recognized both 2-and 1(3)-isomers of monoarachidonoylglycerol with similar efficiencies. The apparent K m and V max values for [$H]2-AG hydrolysis were 91 µM and 29 µM and 2.4 and 1.8 nmol:min −" :mg of protein −" respectively in N18TG2

show abstract

Section: Resultsmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

The novel endogenous cannabinoid 2-arachidonoylglycerol is inactivated by neuronal- and basophil-like cells: connections with anandamide

Marzo

Bisogno

Sugiura

et al. 1998

Biochemical Journal

191

View full text Add to dashboard Cite

show abstract

“…Also anandamide synthase activity was measured in mouse uterus, and was found to respond to sex hormones in the same way as FAAH. Although it is still under debate whether or not anandamide hydrolase and synthase activities belong to the same [22,32] or different [9,18] enzymes, these data demonstrate that the two activities are under the same hormonal control.…”

Section: Discussionmentioning

confidence: 84%

Down-regulation of anandamide hydrolase in mouse uterus by sex hormones

Maccarrone¹,

Felici²,

Bari³

et al. 2000

Eur J Biochem

View full text Add to dashboard Cite

Endocannabinoids are an emerging class of lipid mediators, which mimic several effects of cannabinoids. Anandamide (arachidonoylethanolamide) is a major endocannabinoid, which has been shown to impair pregnancy and embryo development. The activity of anandamide is controlled by cellular uptake through a specific transporter and intracellular degradation by the enzyme anandamide hydrolase (fatty acid amide hydrolase, FAAH). We characterized FAAH in mouse uterus by radiochromatographic and immunochemical techniques, showing that the enzyme is confined to the epithelium and its activity decreases appreciably during pregnancy or pseudopregnancy because of lower gene expression at the translational level. Ovariectomy prevented the decrease in FAAH, and both progesterone and estrogen further reduced its basal levels, suggesting hormonal control of the enzyme. Anandamide was shown to induce programmed cell death in mouse blastocysts, through a pathway independent of type-1 cannabinoid receptor. Blastocysts, however, have a specific anandamide transporter and FAAH, which scavenge this lipid. Taken together, these results provide evidence of an interplay between endocannabinoids and sex hormones in pregnancy. These findings may also be relevant for human fertility, as epithelial cells from healthy human uterus showed FAAH activity and expression, which in adenocarcinoma cells was increased fivefold.Keywords: anandamide hydrolase; apoptosis; development; endocannabinoids; sex hormones.Endocannabinoids are an emerging class of lipid mediators, isolated from brain and peripheral tissues [1,2] and found also in chocolate [3] and milk [4]. They mimic the psychotropic, hypnotic, tranquilizing, antiemetic, anticonvulsive and analgesic effects of cannabinoids [5,6]. These compounds, in particular D 9 -tetrahydrocannabinol, have been reported to have adverse effects on reproductive functions, including retarded embryo development, fetal loss and pregnancy failure [7,8]. A major endocannabinoid, anandamide (arachidonoylethanolamide), has been shown to impair pregnancy and embryo development [9]. Down-regulation of anandamide levels in mouse uterus has been associated with uterine receptivity, while up-regulation correlated with uterine refractoriness to embryo implantation [10]. Remarkably, mouse uterus contains the highest levels of anandamide detected so far in mammalian tissues, and is the only tissue in which anandamide is the main component (up to 95%) of N-acylethanolamines [10]. Anandamide is an endogenous ligand for both the brain-type (CB1-R) and spleen-type (CB2-R) cannabinoid receptors, mimicking several actions of cannabinoids on the central nervous system and in peripheral tissues [11]. Mouse embryos express both CB1-R and CB2-R mRNA, the levels of the former being much higher than those found in brain [7,9,10]. CB1-R activation is detrimental for mouse preimplantation development [10,12], but appears to accelerate trophoblast differentiation [13].The effect of anandamide through CB1-R and CB2-R depends on its concent...

show abstract

“…The resultant supernatant was used as the cytosolic fraction, whereas the pellet was suspended in 20 mM Tris-HCl (pH 7.4) and used as the membrane fraction. Recombinant rat NAPE-PLD highly purified from Escherichia coli (14), the membrane fraction of COS-7 cells overexpressing rat fatty acid amide hydrolase (39), and group IB secretory phospholipase A 2 purified from rat stomach (40) were prepared as described previously.…”

Section: Methodsmentioning

confidence: 99%

Discovery and Characterization of a Ca2+-independent Phosphatidylethanolamine N-Acyltransferase Generating the Anandamide Precursor and Its Congeners

Jin

Okamoto

Morishita

et al. 2007

Journal of Biological Chemistry

Self Cite

137

105

View full text Add to dashboard Cite

N-Acylphosphatidylethanolamines (NAPEs) are precursors of bioactive N-acylethanolamines, including the endocannabinoid anandamide. In animal tissues, NAPE is formed by transfer of a fatty acyl chain at the sn-1 position of glycerophospholipids to the amino group of phosphatidylethanolamine (PE), and this reaction is believed to be the principal rate-limiting step in N-acylethanolamine synthesis. However, the Ca 2؉ -dependent, membrane-associated N-acyltransferase (NAT) responsible for this reaction has not yet been cloned. In this study, on the basis of the functional similarity of NAT to lecithin-retinol acyltransferase (LRAT), we examined a possible PE N-acylation activity in two rat LRAT homologous proteins. Upon overexpression in COS-7 cells, one protein, named rat LRAT-like protein (RLP)-1, catalyzed transfer of a radioactive acyl group from phosphatidylcholine (PC) to PE, resulting in the formation of radioactive NAPE. However, the RLP-1 activity was detected mainly in the cytosolic rather than membrane fraction and was little stimulated by Ca 2؉ . Moreover, RLP-1 did not show selectivity with respect to the sn-1 and sn-2 positions of PC as an acyl donor and therefore could generate N-arachidonoyl-PE (anandamide precursor) from 2-arachidonoyl-PC and PE. In contrast, under the same assay conditions, partially purified NAT from rat brain was highly Ca 2؉ -dependent, membrane-associated, and specific for the sn-1-acyl group of PC. RLP-1 mRNA was expressed predominantly in testis among various rat tissues, and the testis cytosol exhibited an RLP-1-like activity. These results reveal that RLP-1 can function as a PE N-acyltransferase, catalytically distinguishable from the known Ca 2؉ -dependent NAT.

show abstract

Reversible Hydrolysis and Synthesis of Anandamide Demonstrated by Recombinant Rat Fatty-Acid Amide Hydrolase

Cited by 93 publications

References 17 publications

The novel endogenous cannabinoid 2-arachidonoylglycerol is inactivated by neuronal- and basophil-like cells: connections with anandamide

The novel endogenous cannabinoid 2-arachidonoylglycerol is inactivated by neuronal- and basophil-like cells: connections with anandamide

Down-regulation of anandamide hydrolase in mouse uterus by sex hormones

Discovery and Characterization of a Ca2+-independent Phosphatidylethanolamine N-Acyltransferase Generating the Anandamide Precursor and Its Congeners

Contact Info

Product

Resources

About