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An endocannabinoid catabolic enzyme fAAH and its paralogs in an early land plant reveal evolutionary and functional relationship with eukaryotic orthologs imdadul Haq & Aruna Kilaru * endocannabinoids were known to exist only among Animalia but recent report of their occurrence in early land plants prompted us to study its function and metabolism. in mammals, anandamide, as an endocannabinoid ligand, mediates several neurological and physiological processes, which are terminated by fatty acid amide hydrolase (FAAH). We identified nine orthologs of FAAH in the moss Physcomitrella patens (PpFAAH1 to PpFAAH9) with amidase signature and catalytic triad. The optimal amidase activity for PpFAAH1 was at 37 °C and pH 8.0, with higher specificity to anandamide. Further, the phylogeny and predicted structural analyses of the nine paralogs revealed that PpFAAH1 to PpFAAH4 were closely related to plant FAAH while PpFAAH6 to PpFAAH9 were to the rat FAAH, categorized based on the membrane binding cap, membrane access channel and substrate binding pocket. We also identified that a true 'dynamic paddle' that is responsible for tighter regulation of FAAH is recent in vertebrates and absent or not fully emerged in plants and non-vertebrates. these data reveal evolutionary and functional relationship among eukaryotic fAAH orthologs and features that contribute to versatility and tighter regulation of fAAH. future studies will utilize fAAH mutants of moss to elucidate the role of anandamide in early land plants. Endocannabinoids such as anandamide belong to a class of lipid derivatives referred to as N-acylethalomines (NAEs). These fatty acid ethanolamides are found in wide range of eukaryotic organisms such as yeast, Caenorhabditis elegans, bivalve mollusk, mammals and also plants 1-4. The fatty acid chain length of these NAEs can range from 12 C to 20 C and are either saturated or unsaturated 5. Among these NAEs, only anandamide or N-arachidonylethanolamide (NAE 20:4) is known to serve as a ligand to cannabinoid binding receptors to activate endocannabinoid signaling 6. In mammals, while endocannabinoids are key participants in neural signaling and other physiological events 7-11 , other NAEs such as NAE16:0 serve protective function in a receptor-independent manner 12. Irrespective of their receptor binding capability, the NAE type, content, composition and functions are highly diverse among organisms, in a tissue-specific manner. In C. elegans, endocannabinoids, including anandamide are essential for mobilization of cholesterol from internal reserves 13 and they affect life span 14. In human, circulating endocannabinoids are positively correlated to visceral adipose tissue mass 15 , while in rat, anandamide induces overeating by activating cannabinoid receptor 16. In higher plants such as Arabidopsis thaliana, only 12 C to 18 C NAEs occur and they mediate growth, development and biotic and abiotic stress responses 17-21. Interestingly, the fatty acid and NAE composition of early land plants such as bryophytes is distinct from th...
An endocannabinoid catabolic enzyme fAAH and its paralogs in an early land plant reveal evolutionary and functional relationship with eukaryotic orthologs imdadul Haq & Aruna Kilaru * endocannabinoids were known to exist only among Animalia but recent report of their occurrence in early land plants prompted us to study its function and metabolism. in mammals, anandamide, as an endocannabinoid ligand, mediates several neurological and physiological processes, which are terminated by fatty acid amide hydrolase (FAAH). We identified nine orthologs of FAAH in the moss Physcomitrella patens (PpFAAH1 to PpFAAH9) with amidase signature and catalytic triad. The optimal amidase activity for PpFAAH1 was at 37 °C and pH 8.0, with higher specificity to anandamide. Further, the phylogeny and predicted structural analyses of the nine paralogs revealed that PpFAAH1 to PpFAAH4 were closely related to plant FAAH while PpFAAH6 to PpFAAH9 were to the rat FAAH, categorized based on the membrane binding cap, membrane access channel and substrate binding pocket. We also identified that a true 'dynamic paddle' that is responsible for tighter regulation of FAAH is recent in vertebrates and absent or not fully emerged in plants and non-vertebrates. these data reveal evolutionary and functional relationship among eukaryotic fAAH orthologs and features that contribute to versatility and tighter regulation of fAAH. future studies will utilize fAAH mutants of moss to elucidate the role of anandamide in early land plants. Endocannabinoids such as anandamide belong to a class of lipid derivatives referred to as N-acylethalomines (NAEs). These fatty acid ethanolamides are found in wide range of eukaryotic organisms such as yeast, Caenorhabditis elegans, bivalve mollusk, mammals and also plants 1-4. The fatty acid chain length of these NAEs can range from 12 C to 20 C and are either saturated or unsaturated 5. Among these NAEs, only anandamide or N-arachidonylethanolamide (NAE 20:4) is known to serve as a ligand to cannabinoid binding receptors to activate endocannabinoid signaling 6. In mammals, while endocannabinoids are key participants in neural signaling and other physiological events 7-11 , other NAEs such as NAE16:0 serve protective function in a receptor-independent manner 12. Irrespective of their receptor binding capability, the NAE type, content, composition and functions are highly diverse among organisms, in a tissue-specific manner. In C. elegans, endocannabinoids, including anandamide are essential for mobilization of cholesterol from internal reserves 13 and they affect life span 14. In human, circulating endocannabinoids are positively correlated to visceral adipose tissue mass 15 , while in rat, anandamide induces overeating by activating cannabinoid receptor 16. In higher plants such as Arabidopsis thaliana, only 12 C to 18 C NAEs occur and they mediate growth, development and biotic and abiotic stress responses 17-21. Interestingly, the fatty acid and NAE composition of early land plants such as bryophytes is distinct from th...
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