Identification and recombinant expression of anandamide hydrolyzing enzyme from Dictyostelium discoideum

  1. Get@NRC: Identification and recombinant expression of anandamide hydrolyzing enzyme from Dictyostelium discoideum (Opens in a new window)
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Journal titleBMC Microbiology
Article number124
Subjectanandamide; epitope; fatty acid amidase; hybrid protein; article; catalysis; Dictyostelium discoideum; DNA sequence; Escherichia coli; fruiting body; in vivo study; life cycle; nonhuman; nucleotide sequence; protein expression; protein localization; sequence analysis; Amidohydrolases; Amino Acid Sequence; Arachidonic Acids; Cloning, Molecular; Computational Biology; Dictyostelium; DNA, Complementary; Endocannabinoids; Escherichia coli; Gene Expression; Gene Expression Profiling; Membrane Proteins; Molecular Sequence Data; Polyunsaturated Alkamides; Recombinant Proteins; Sequence Alignment; Sequence Homology, Amino Acid; Substrate Specificity; Dictyostelium; Dictyostelium discoideum; Eukaryota
AbstractBackground: Anandamide (Arachidonoyl ethanolamide) is a potent bioactive lipid studied extensively in humans, which regulates several neurobehavioral processes including pain, feeding and memory. Bioactivity is terminated when hydrolyzed into free arachidonic acid and ethanolamine by the enzyme fatty acid amide hydrolase (FAAH). In this study we report the identification of a FAAH homolog from Dictyostelium discoideum and its function to hydrolyze anandamide. Results: A putative FAAH DNA sequence coding for a conserved amidase signature motif was identified in the Dictyostelium genome database and the corresponding cDNA was isolated and expressed as an epitope tagged fusion protein in either E.coli or Dictyostelium. Wild type Dictyostelium cells express FAAH throughout their development life cycle and the protein was found to be predominantly membrane associated. Production of recombinant HIS tagged FAAH protein was not supported in E.coli host, but homologous Dictyostelium host was able to produce the same successfully. Recombinant FAAH protein isolated from Dictyostelium was shown to hydrolyze anandamide and related synthetic fatty acid amide substrates. Conclusions: This study describes the first identification and characterisation of an anandamide hydrolyzing enzyme from Dictyostelium discoideum, suggesting the potential of Dictyostelium as a simple eukaryotic model system for studying mechanisms of action of any FAAH inhibitors as drug targets. © 2012 Monecke et al.; licensee BioMed Central Ltd.
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AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Biological Sciences (IBS-ISB)
Peer reviewedYes
NPARC number21269328
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Record identifier50e627c0-7c70-478a-9867-25c1676a7c6e
Record created2013-12-12
Record modified2016-05-09
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