Two novel classes of neuroactive fatty acid amides are substrates for mouse neuroblastoma 'anandamide amidohydrolase'
| Autor: | Vincenzo Di Marzo, Aldo Di Luccia, Tiziana Bisogno, Gennaro Marino, S. Maurelli, Luciano De Petrocellis |
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| Rok vydání: | 1995 |
| Předmět: |
Hot Temperature
Stereochemistry Polyunsaturated Alkamides medicine.medical_treatment Biophysics oleoyl-amide Oleic Acids Arachidonic Acids Biochemistry Binding Competitive Amidohydrolases Substrate Specificity Hydrolysis chemistry.chemical_compound Mice Neuroblastoma Structural Biology Enzyme Stability Genetics medicine Tumor Cells Cultured anandamide fatty acid amide Peptide bond Animals Molecular Biology chemistry.chemical_classification Fatty acid amide Fatty Acids Fatty acid Cell Biology Anandamide arachidonoyl-ethanolamide cannabinoid Hydrogen-Ion Concentration Sleep-inducing factor Endocannabinoid system Amides Enzyme chemistry Cannabinoid Endocannabinoids Oleic Acid |
| Zdroj: | FEBS letters 377 (1995): 82–86. doi:10.1016/0014-5793(95)01311-3 info:cnr-pdr/source/autori:Maurelli S, Bisogno T, De Petrocellis L, Di Luccia A, Marino G, Di Marzo V./titolo:Two novel classes of neuroactive fatty acid amides are substrates for mouse neuroblastoma 'anandamide amidohydrolase'./doi:10.1016%2F0014-5793(95)01311-3/rivista:FEBS letters (Print)/anno:1995/pagina_da:82/pagina_a:86/intervallo_pagine:82–86/volume:377 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/0014-5793(95)01311-3 |
| Popis: | The endogenous cannabimimetic substance, anandamide (N-arachidonoyl-ethanolamine) and the recently isolated sleep-inducing factor, oleoyl-amide (cis-9,10-octadecenoamide), belong to two neuroactive fatty acid amide classes whose action in mammals has been shown to be controlled by enzymatic amide bond hydrolysis. Here me report the partial characterisation and purification of 'anandamide amidohydrolase' from membrane fractions of N18 neuroblastoma cells, and provide evidence for a further and previously unsuspected role of this enzyme, An enzymatic activity catalysing the hydrolysis of [C-14]anandamide was found in both microsomal and 10,000 x g pellet fractions, The latter fractions, which displayed the highest V-max for anandamide, were used for further characterisation of the enzyme, and were found to catalyse the hydrolysis also of [C-14]oleoyl-amide, with an apparent K-m of 9.0 +/- 2.2 mu M. [C-14]anandamide- and [C-14]oleoyl-amide-hydrolysing activities: (i) exhibited identical pH- and temperature-dependency profiles; (ii) were inhibited by alkylating agents; (iii) were competitively inhibited by the phospholipase A(2) inhibitor arachidonyl-trifluoromethyl-ketone with the same IC50 (3 mu M); (iv) were competitively inhibited by both anandamide (or other polyunsaturated fatty acid-ethanolamides) and oleoyl-amide, Proteins solubilised from 10,000 x g pellets were directly analysed by isoelectric focusing, yielding purified fractions capable of catalysing the hydrolysis of both [C-14]anandamide and [C-14]oleoyl-amide. These data suggest that 'anandamide amidohydrolase' enzymes, such as that characterised in this study, may be used by neuronal cells also to hydrolyse the novel sleep-inducing factor oleoyl-amide. |
| Databáze: | OpenAIRE |
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