title: New fluorescent probes for ligand-binding assays of odorant-binding proteins.
abstract: Fluorescence-linked binding assays allow determination of dissociation constants at equilibrium and have recently become increasingly popular, thanks to their ease of operation. Currently used probes, such as 1-aminoanthracene and N-phenyl-1-naphthylamine, are excited and emit in the ultraviolet region, but alternative ligands operating in the visible spectrum would be highly desirable for applications in biosensing devices. Based on the two above structures, we have designed and synthesised six new fluorescent probes to be used in ligand-binding assays. The compounds are derivatives of naphatalene, anthracene and fluoranthene and present two aromatic moieties linked by an amine nitrogen. We have measured the emission spectra of the new probes and their binding to three odorant-binding proteins. The probes bind the tested proteins with different affinities, generally with dissociation constants about one order of magnitude lower than the parent compounds. The extended aromatic systems present in the new compounds produced a shift of both excitation and emission peaks at higher wavelength, close or within the visible spectrum, thus facilitating measurements in biosensors for odorants and small organic molecules using optical devices.
authors.affiliation: Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy.
authors.forename: Rosa
authors.initials: R
authors.lastname: Mastrogiacomo
authors.affiliation: Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy.
authors.forename: Immacolata
authors.initials: I
authors.lastname: Iovinella
authors.affiliation: Scuola Normale Superiore and Department of Pharmacy of the University of Pisa, Pisa, Italy. Electronic address: elio.napolitano@farm.unipi.it.
authors.forename: Elio
authors.initials: E
authors.lastname: Napolitano
mesh_terms: D000818:Animals; D001516:Bees; D015374:Biosensing Techniques; D012831:Bombyx; D015195:Drug Design; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D019476:Insect Proteins / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D015394:Molecular Structure; D011485:Protein Binding; D018035:Receptors, Odorant / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine
chemical_list: D005456:Fluorescent Dyes; D019476:Insect Proteins; D008024:Ligands; D018035:Receptors, Odorant; D011994:Recombinant Proteins; C045648:odorant-binding protein
keywords: Anthracene derivatives; Fluoranthene derivatives; Fluorescent probe; Ligand-binding assay; Naphtalene derivatives; Odorant-binding proteins