Dioxygenation of N-linoleoyl amides by soybean lipoxygenase-1

• Published in: FEBS letters 1997-07, Vol.411 (2), p.287-290
• Main Authors: van der Stelt, Marcelis, Nieuwenhuizen, Willem F, Veldink, Gerrit A, Vliegenthart, Johannes F.G
• Format: Article
• Language: English
• Subjects: 13-HOD, 13-hydroxy-9Z,11E-octadeca-9,11-dienoic acid; 13-HODN(Me)2, 13-hydroxy-9Z,11E-octadeca-9,11-dienoyl-N,N-dimethylamide; 13-HODNH2, 13-hydroxy-9Z,11E-octadeca-9,11-dienoyl-N-amide; 13-HODNHEtOH, 13-hydroxy-9Z,11E-octadeca-9,11-dienoyl-N-ethanolamide; 13-HODNHMe, 13-hydroxy-9Z,11E-octadeca-9,11-dienoyl-N-methylamide; 1H-NMR, 1H-nuclear magnetic resonance; 2D-NMR, 2-dimensional nuclear magnetic resonance; Amides - chemistry; Amides - metabolism; Arachidonic Acids - metabolism; Cannabinoid; CB1/2, cannabinoid binding receptor 1 or 2; CD, circular dichroism; CP-HPLC, chiral-phase high-performance liquid chromatography; Endocannabinoids; Fatty acid amide; Fatty Acids - metabolism; FPLC, fast protein liquid chromatography; FT-IR, Fourier transform infrared (spectrometry); GC-MS, gas chromatography-mass spectrometry; Glycine max - enzymology; Kinetics; Linoleic Acids - metabolism; Lipoxygenase; Lipoxygenase - metabolism; LOX-1, lipoxygenase-1; Magnetic Resonance Spectroscopy; Molecular Conformation; Molecular Structure; N-Linoleoyl amide; N-Linoleoyl ethanolamide; Neurotransmitter Agents - metabolism; OD, 9Z,12Z-octadecadienoicacid (linoleic acid); ODN(Me)2, 9Z,12Z-octadeca-9,12-dienoyl-N,N-dimethylamide; ODNH2, 9Z,12Z-octadeca-9,12-dienoyl-N-amide; ODNHEtOH, 9Z,12Z-octadeca-9,12-dienoyl-N-ethanolamide; ODNHMe, 9Z,12Z-octadeca-9,12-dienoyl-N-methylamide; Polyunsaturated Alkamides; RP-HPLC, reversed-phase high-performance liquid chromatography; Spectrophotometry; THC, tetrahydrocannabinol; TOCSY, total correlation spectroscopy; UV, ultraviolet (spectrophotometry)
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)00718-7

Anandamide, a novel neurotransmitter, has been reported to be dioxygenated by brain lipoxygenase [1, 11]. Anandamides constitute a new class of neuroregulatory fatty acid amides. However, little is known about the enzymatic dioxygenation of these lipids. Therefore, we have tested several members of the neuroactive fatty acid amide class containing a 1 Z,4 Z-pentadiene system whether they could be dioxygenated by soybean lipoxygenase-1, which is a model enzyme for mammalian lipoxygenases. In this study it was found that lipoxygenase-1 converts N-linoleoylethanolamide (ODNHEtOH), N-linoleoylamide (ODNH 2), N-linoleoylmethylamide (ODNHMe) and N, N-linoleoyldimethylamide (ODN(Me) 2 into 13-( S)-hydroperoxy-9 Z,11 E-octadeca-9,11-dienoyl amides derivatives. The apparent K m values for ODNHEtOH (23.6±3.7 μM), ODNH 2 (8.60±0.65 μM) and linoleic acid (OD: 8.85±0.74 μM) are not significantly different. The k cat for ODNH 2 (32.4±1.2 s −1) is twice as small as compared to the turnover numbers of the other substrates, viz. ODNHEtOH (61.6±5.0 s −1) and OD (54.4±2.0 s −1). The results suggest that N-linoleoyl ethanolamide and N-linoleoyl amide can be readily converted by lipoxygenases in vivo.