Pharmacological folding chaperones act as allosteric ligands of Frizzled4.
Upon binding, ligands can chaperone their protein targets by preventing them from misfolding and aggregating. Thus, an organic molecule that works as folding chaperone for a protein might be its specific ligand, and, similarly, the chaperone potential could represent an alternative readout in a molecular screening campaign toward the identification of new hits. Here we show that small molecules selected for acting as pharmacological chaperones on a misfolded mutant of the Frizzled4 (Fz4) receptor bind and modulate wild-type Fz4, representing what are to our knowledge the first organic ligands of this until-now-undruggable GPCR. The novelty and the advantages of the screening platform, the allosteric binding site addressed by these new ligands and the mechanism they use to modulate Fz4 suggest new avenues for development of inhibitors of the Wnt-β-catenin pathway and for drug discovery.
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
Serena F
SF
Generoso
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Mariateresa
M
Giustiniano
Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza University of Rome, Rome, Italy.
Giuseppe
G
La Regina
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Sara
S
Bottone
Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza University of Rome, Rome, Italy.
Sara
0000000197810621
S
Passacantilli
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Salvatore
S
Di Maro
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Hilde
H
Cassese
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Agostino
A
Bruno
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
Massimo
M
Mallardo
Department of Clinical Medicine and Surgery, University of Naples Federico II, Naples, Italy.
Monica
M
Dentice
Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza University of Rome, Rome, Italy.
Romano
R
Silvestri
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Luciana
L
Marinelli
1] Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy. [2] CEINGE-Biotecnologie Avanzate, s.c.a.r.l., Naples, Italy.
Daniela
D
Sarnataro
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
Stefano
S
Bonatti
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Ettore
E
Novellino
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Mariano
M
Stornaiuolo
D000495:Allosteric Site; D020816:Amino Acid Motifs; D001483:Base Sequence; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D051157:Frizzled Receptors / Q000737:chemistry*; D005990:Glycerol / Q000737:chemistry; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008856:Microscopy, Fluorescence; D018832:Molecular Chaperones / Q000737:chemistry*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011485:Protein Binding; D017510:Protein Folding; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry
C402930:FZD4 protein, human; D051157:Frizzled Receptors; D008024:Ligands; D018832:Molecular Chaperones; D043562:Receptors, G-Protein-Coupled; D005990:Glycerol