title: Fluorine nuclear magnetic resonance-based assay in living mammalian cells.
abstract: Nuclear magnetic resonance (NMR)-based screening has been recognized as a powerful approach for the identification and characterization of molecules interacting with pharmaceutical targets. Indeed, several NMR methods have been developed and successfully applied to many drug discovery projects. Whereas most of these approaches have targeted isolated biomolecular receptors, very few cases are reported with the screening performed in intact cells and cell extracts. Here we report the first successful application of the fluorine NMR-based assay n-FABS (n-fluorine atoms for biochemical screening) in living mammalian cells expressing the membrane protein fatty acid amide hydrolase (FAAH). This method allows the identification of both weak and potent inhibitors and the measurement of their potency in a physiological environment.
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy. Electronic address: marina.veronesi@iit.it.
authors.forename: Marina
authors.initials: M
authors.lastname: Veronesi
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Giacomina
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Romeo
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Castellani
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Ottonello
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Lambruschini
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Gianpiero
authors.initials: G
authors.lastname: Garau
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Rita
authors.initials: R
authors.lastname: Scarpelli
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy.
authors.forename: Tiziano
authors.initials: T
authors.lastname: Bandiera
authors.affiliation: Department of Drug Discovery and Development, Fondazione Istituto Italiano di Tecnologia, 16163 Genoa, Italy; Department of Anatomy and Neurobiology, School of Medicine, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Daniele
authors.initials: D
authors.lastname: Piomelli
authors.affiliation: Institut de Chimie, Faculté des Sciences, Université de Neuchâtel, 2000 Neuchâtel, Switzerland.
authors.forename: Claudio
authors.initials: C
authors.lastname: Dalvit
mesh_terms: D000581:Amidohydrolases / Q000032:analysis* / Q000378:metabolism; D001549:Benzamides / Q000737:chemistry / Q000378:metabolism; D002219:Carbamates / Q000737:chemistry / Q000378:metabolism; D005461:Fluorine / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D019906:Nuclear Magnetic Resonance, Biomolecular; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D001549:Benzamides; D002219:Carbamates; D005461:Fluorine; D000099252:Fatty Acid Amide Hydrolases; C500528:cyclohexyl carbamic acid 3'-carbamoylbiphenyl-3-yl ester
keywords: (19)F NMR screening; IC(50); Mammalian cells; Metabolic fingerprint; hFAAH; n-FABS