title: Novel fluorescent cycloheximide derivatives for the imaging of protein synthesis.
abstract: Cycloheximide (CHX) is one of the most interesting protein synthesis inhibitors. For this reason, fluorescent derivatives of CHX could find useful applications in cell biology. We report the successful synthesis of a set of novel fluorescent derivatives of CHX. The effect of different functional groups on the biological activity of CHX was studied upon their modification through suitable strategies, i.e., acetylation of the hydroxyl group and reductive amination of the ketone group. The first route induced a complete loss of biological activity, while the second approach allowed a retained inhibition of protein synthesis, as demonstrated by in vitro translation assays. Various fluorescent dyes for reductive amination were tested (i.e., ANTS, APTS, and Rhodamine-123), and the success of the syntheses was demonstrated by diverse analytical techniques. Cycloheximide labeling with fluorescent dyes is a promising approach for developing fluorescence reporters for various applications, both in vitro (fluorescence spectroscopy) and in vivo (live imaging).
authors.affiliation: Neurobiology Sector, SISSA-ISAS, Trieste, Italy. f.paoletti@ebri.it
authors.forename: Francesca
authors.initials: F
authors.lastname: Paoletti
authors.forename: Kevin
authors.initials: K
authors.lastname: Ainger
authors.forename: Ivan
authors.initials: I
authors.lastname: Donati
authors.forename: Raffaella
authors.initials: R
authors.lastname: Scardigli
authors.forename: Amedeo
authors.initials: A
authors.lastname: Vetere
authors.forename: Antonino
authors.initials: A
authors.lastname: Cattaneo
authors.forename: Cristiana
authors.initials: C
authors.lastname: Campa
mesh_terms: D000107:Acetylation; D000818:Animals; D003513:Cycloheximide / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D051379:Mice; D041681:NIH 3T3 Cells; D014176:Protein Biosynthesis; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D003513:Cycloheximide