The Synthesis of Eicosanoyl-5-hydroxytryptamide in Mammalian Tissue

Abstract

Alzheimer’s disease (AD) and Parkinson’s disease (PD) are the two most common forms of neurodegeneration, creating a large health concern and a popular focus for neurological research. Recent studies have shown decreased risk for both diseases associated with coffee consumption. In a scan of coffee compounds, eicosanoyl-5- hyproxytryptamide (EHT) was identified and later shown to be neuroprotective in rat and mouse models of AD and PD respectively. Furthermore, EHT is likely to be synthesized in mammalian tissue, given recent findings of N-acyl serotonin synthesis in the porcine gastro-intestinal tract. The responsible enzyme, EHT synthase, has not yet been identified. Due to the promise of EHT as a potential therapeutic target against neurodegeneration, this study seeks to understand its synthesis in mammalian tissue. We begin by examining EHT formation in bovine brain, using radioactive tracing of substrates and thin-layer chromatography analysis, yielding inconclusive results. High performance liquid chromatography analysis was also performed while analyzing EHT formation in rat tissues, providing evidence for EHT synthesis in rat intestinal tissue, where EHT synthase is likely membrane-bound. Finally, bioinformatic analysis was used to analytically identify candidate sequences for EHT synthase based on its likely relationship with other known and characterized enzymes, arylalkylamine Nacetyltransferase (AANAT) and ceramide synthase (CerS). The findings presented here strongly suggest that EHT is synthesized in the human body. A full understanding of EHT synthesis could lead to the ability to regulate its formation, making it a viable drug target in the prevention of neurodegenerative disease.