Equilibrium in the hydrolysis and synthesis of cannabimimetic anandamide demonstrated by a purified enzyme

Katayama, Kazuhisa; Ueda, Natsuo; Katoh, Itsuo; Yamamoto, Shozo

doi:10.1016/s1388-1981(99)00124-9

Cited by 56 publications

(39 citation statements)

References 32 publications

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“…N-Palmitoylethanolamine was hydrolyzed 13 times faster than anandamide. This substrate specificity was in contrast with that of anandamide amidohydrolase, which prefers anandamide to N-palmitoylethanolamine (9,28,29,34,35). Thus, we refer to the enzyme with higher reactivity for N-palmitoylethanolamine as N-palmitoylethanolamine hydrolase.…”

Section: Discussionmentioning

confidence: 94%

“…The enzyme is widely distributed in mammalian organs (28,30,31) and characterized by an optimal pH value at 8.5-10 (9, 29) and by high sensitivity to phenylmethylsulfonyl fluoride (27) and methyl arachidonyl fluorophosphonate (MAFP) 1 (32,33). The enzyme has a broad substrate specificity but hydrolyzes N-palmitoylethanolamine at a lower rate than anandamide (9,28,29,34,35).…”

mentioning

confidence: 99%

“…Solubilization of the enzyme from the 12,000 ϫ g pellet was performed as described above. Recombinant rat anandamide amidohydrolase was overexpressed in a baculovirusSf9 insect cell system (35), and the particulate fraction of the transfected Sf9 cells was used for enzyme assay.…”

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confidence: 99%

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Purification and Characterization of an Acid Amidase Selective for N-Palmitoylethanolamine, a Putative Endogenous Anti-inflammatory Substance

Ueda¹,

Yamanaka²,

Yamamoto³

2001

Journal of Biological Chemistry

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231

238

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N-Arachidonoylethanolamine (anandamide) is cannabimimetic, and N-palmitoylethanolamine is anti-inflammatory and immunosuppressive. We found an amidase that is more active with the latter than the former in contrast to the previously known anandamide amidohydrolase for which N-palmitoylethanolamine is a poor substrate. Proteins solubilized by freezing and thawing from the 12,000 ؋ g pellet of various rat organs hydrolyzed [ 14 C]N-palmitoylethanolamine to palmitic acid and ethanolamine. The specific enzyme activity was higher in the order of lung > spleen > small intestine > thymus > cecum, and high activity was found in peritoneal and alveolar macrophages. The enzyme with a molecular mass of 31 kDa was purified from rat lung to a specific activity of 1.8 mol/min/mg protein. Relative reactivities of the enzyme with various N-acylethanolamines (100 M) were as follows: N-palmitoylethanolamine, 100%; N-myristoylethanolamine, 48%; N-stearoylethanolamine, 21%; N-oleoylethanolamine, 20%; N-linoleoylethanolamine, 13%; anandamide, 8%. The enzyme was the most active at pH 5 and was activated 7-fold by Triton X-100. The enzyme was almost insensitive to methyl arachidonyl fluorophosphonate, which inhibited anandamide amidohydrolase potently. Thus, the new enzyme referred to as N-palmitoylethanolamine hydrolase was clearly distinguishable from anandamide amidohydrolase.

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Section: Discussionmentioning

confidence: 94%

mentioning

confidence: 99%

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Purification and Characterization of an Acid Amidase Selective for N-Palmitoylethanolamine, a Putative Endogenous Anti-inflammatory Substance

Ueda¹,

Yamanaka²,

Yamamoto³

2001

Journal of Biological Chemistry

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231

238

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“…1). Since cells lacking NAPEphospholipase D are also able to synthesise anandamide, alternative pathways have been proposed (41)(42)(43)(44)(45)(46)(47) and they are summarised in Fig. 1.…”

Section: Endocannabinoids Biosynthesis and Uptakementioning

confidence: 99%

PUFA-derived endocannabinoids: an overview

Cascio

2013

Proc. Nutr. Soc.

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Following on from the discovery of cannabinoid receptors, of their endogenous agonists (endocannabinoids) and of the biosynthetic and metabolic enzymes of the endocannabinoids, significant progress has been made towards the understanding of the role of the endocannabinoid system in both physiological and pathological conditions. Endocannabinoids are mainly n-6 long-chain PUFA (LCPUFA) derivatives that are synthesised by neuronal cells and inactivated via a two-step process that begins with their transport from the extracellular to the intracellular space and culminates in their intracellular degradation by hydrolysis or oxidation. Although the enzymes responsible for the biosynthesis and metabolism of endocannabinoids have been well characterised, the processes involved in their cellular uptake are still a subject of debate. Moreover, little is yet known about the roles of endocannabinoids derived from n-3 LCPUFA such as EPA and DHA. Here, I provide an overview of what is currently known about the mechanisms for the biosynthesis and inactivation of endocannabinoids, together with a brief analysis of the involvement of the endocannabinoids in both food intake and obesity. Owing to limited space, recent reviews will be often cited instead of original papers.

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“…This is so because this reaction requires catalysis by the fatty acid amide hydrolase (FAAH) working in reverse. In vitro, this, in turn, requires very high concentrations of arachidonic acid and ethanolamine to drive the enzyme to anandamide production instead of hydrolysis (12)(13)(14). The more natural reaction catalyzed by FAAH (i.e., the hydrolysis of anandamide to yield arachidonoic acid and ethanolamine instead) occurs at more physiological concentrations of substrate and products.…”

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confidence: 99%