title: Luciferin Amides Enable in Vivo Bioluminescence Detection of Endogenous Fatty Acid Amide Hydrolase Activity.
abstract: Firefly luciferase is homologous to fatty acyl-CoA synthetases. We hypothesized that the firefly luciferase substrate d-luciferin and its analogs are fatty acid mimics that are ideally suited to probe the chemistry of enzymes that release fatty acid products. Here, we synthesized luciferin amides and found that these molecules are hydrolyzed to substrates for firefly luciferase by the enzyme fatty acid amide hydrolase (FAAH). In the presence of luciferase, these molecules enable highly sensitive and selective bioluminescent detection of FAAH activity in vitro, in live cells, and in vivo. The potency and tissue distribution of FAAH inhibitors can be imaged in live mice, and luciferin amides serve as exemplary reagents for greatly improved bioluminescence imaging in FAAH-expressing tissues such as the brain.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, United States.
authors.forename: David M
authors.initials: DM
authors.lastname: Mofford
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, United States.
authors.forename: Spencer T
authors.initials: ST
authors.lastname: Adams
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, United States.
authors.forename: G S Kiran Kumar
authors.initials: GS
authors.lastname: Reddy
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, United States.
authors.forename: Gadarla Randheer
authors.initials: GR
authors.lastname: Reddy
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation Street, Worcester, Massachusetts 01605, United States.
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Miller
mesh_terms: D000577:Amides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D000581:Amidohydrolases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D052160:Benzothiazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D016466:CHO Cells; D003412:Cricetulus; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D049409:Luciferases, Firefly / Q000378:metabolism*; D049408:Luminescent Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D061848:Optical Imaging; D010880:Piperidines / Q000493:pharmacokinetics* / Q000494:pharmacology; D011725:Pyridines / Q000493:pharmacokinetics* / Q000494:pharmacology; D014018:Tissue Distribution; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000577:Amides; D000581:Amidohydrolases; D052160:Benzothiazoles; D004791:Enzyme Inhibitors; D049409:Luciferases, Firefly; D049408:Luminescent Agents; D010880:Piperidines; D011725:Pyridines; D000099252:Fatty Acid Amide Hydrolases; C532924:D-luciferin; C557973:PF 3845