Solid-phase organic tagging resins for labeling biomolecules by 1,3-dipolar cycloaddition: application to the synthesis of a fluorescent non-peptidic vasopressin receptor ligand.
Two novel solid-phase organic tagging (SPOrT) resins were synthesized to facilitate the labeling of peptides and small organic compounds with a fluorescent probe. Both resins were obtained from the commercially available backbone amide linker (BAL) resin. Following the solid-phase synthesis of model compounds, a tripeptide and benzazepine, the fluorescent probe derived from Lissamine Rhodamine B was incorporated through CuI-catalyzed 1,3-dipolar cycloaddition. Final cleavage in acidic media enabled access to both types of molecules in good yield with high purity. The SPOrT resin was successfully applied to the preparation of the first non-peptidic fluorescent compound with a nanomolar affinity for the human vasopressin V2 receptor (V2R) subtype. This molecule will find application in binding assays that use polarization or fluorescence resonance energy-transfer (FRET) techniques. The SPOrT resins are also well suited for other tags and the parallel synthesis of a fluorescently tagged library for protein screening.
Département de Pharmacochimie de la Communication Cellulaire, Université Strasbourg 1, Institut Gilbert Laustriat, CNRS UMR 7175, Faculté de Pharmacie, 74 route du Rhin, BP 60024, 67401 Illkirch, France. dominique.bonnet@pharma.u-strasbg.fr
Dominique
D
Bonnet
Stéphanie
S
Riché
Stéphanie
S
Loison
Rania
R
Dagher
Marie-Céline
MC
Frantz
Laure
L
Boudier
Rita
R
Rahmeh
Bernard
B
Mouillac
Jacques
J
Haiech
Marcel
M
Hibert
D004563:Electrochemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D008024:Ligands; D008094:Lithium / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D017483:Receptors, Vasopressin / Q000737:chemistry* / Q000378:metabolism; D013059:Spectrum Analysis, Raman; D014867:Water / Q000737:chemistry
D005456:Fluorescent Dyes; D008024:Ligands; D010455:Peptides; D017483:Receptors, Vasopressin; D014867:Water; D008094:Lithium