N-acetylation of biogenic amines in Ascaridia galli

Abstract

SUMMARYThe metabolism of 5-hydroxytryptamine (5-HT), octopamine (OA) and dopamine (DA) by adult male Ascaridia galli was investigated by incubating worms cut into 5 mm lengths with radio-isotope labelled amine. [14C]5-hydroxytryptamine and [14C]octopamine were taken up by the tissues and were metabolized to apolar products that co-chromatographed using both high-performance liquid chromatography and thin-layer chromatography with N-acetyl 5-hydroxytryptamine (NA5-HT) and N-acetyloctopamine (NAOA), respectively. N-acetylation was by far the most important reaction detected under these experimental conditions. A brief incubation of cut worms with [3H]dopamine resulted in the formation of a radio-isotope labelled metabolite that co-chromatographed with N-acetyldopamine (NADA) on reversed-phase high-performance liquid chromatography. The N-acetyltransferase activity towards 5-hydroxytryptamine and octopamine was detected in crude homogenates of male worms only when the co-substrate, acetyl CoA, was added to the reaction mixture. This enzyme activity appeared to be mainly localized in a 40 000 g supernatant fraction. The failure of previous studies to detect N-acetylation of biogenic amines in tissue homogenates of nematodes may have been due to the low levels of acetyl CoA present in these enzyme preparations.