TY - JOUR
T1 - A conserved molecular switch in Class F receptors regulates receptor activation and pathway selection
AU - Wright, Shane C.
AU - Kozielewicz, Paweł
AU - Kowalski-Jahn, Maria
AU - Petersen, Julian
AU - Bowin, Carl Fredrik
AU - Slodkowicz, Greg
AU - Marti-Solano, Maria
AU - Rodríguez, David
AU - Hot, Belma
AU - Okashah, Najeah
AU - Strakova, Katerina
AU - Valnohova, Jana
AU - Babu, M. Madan
AU - Lambert, Nevin A.
AU - Carlsson, Jens
AU - Schulte, Gunnar
N1 - Funding Information:
We thank Anna Krook for access to the ClarioStar plate reader, Benoit Vanhollebeke for providing the HEK293TΔFZD1-10 cells, and Roger Sunahara for permission to use and modify his receptor and heterotrimer cartoons. The work was supported by grants from Karolinska Institutet, the Swedish Research Council (2013-5708, 2015-02899, 2017-04676), the Swedish Cancer Society (CAN2014/659, CAN2017/561), the Novo Nordisk Foundation (NNF17OC0026940), the Knut & Alice Wallenberg Foundation (KAW2008.0149), Stiftelsen Olle Engkvist Byggmästare (2016/193), Emil and Wera Cornells Stiftelse, the Swedish Society for Medical Research (SSMF), the Science for Life Laboratory, the Marie Curie ITN WntsApp (Grant no. 608180; www.wntsapp.eu) and the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation; grant no. KO 5463/1-1). M.M.S. received support from a FEBS Long-Term Fellowship. Computational resources were provided by the Swedish National Infrastructure for Computing (SNIC), High Performance Computing Centre North (HPC2N) in Umeå, National Supercomputer Centre (NSC) in Linköping, Uppsala Multidisciplinary Center for Advanced Computational Science (UPPMAX), and the Medical Research Council, UK (MC_U105185859).
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Class F receptors are considered valuable therapeutic targets due to their role in human disease, but structural changes accompanying receptor activation remain unexplored. Employing population and cancer genomics data, structural analyses, molecular dynamics simulations, resonance energy transfer-based approaches and mutagenesis, we identify a conserved basic amino acid in TM6 in Class F receptors that acts as a molecular switch to mediate receptor activation. Across all tested Class F receptors (FZD 4,5,6,7, SMO), mutation of the molecular switch confers an increased potency of agonists by stabilizing an active conformation as assessed by engineered mini G proteins as conformational sensors. Disruption of the switch abrogates the functional interaction between FZDs and the phosphoprotein Dishevelled, supporting conformational selection as a prerequisite for functional selectivity. Our studies reveal the molecular basis of a common activation mechanism conserved in all Class F receptors, which facilitates assay development and future discovery of Class F receptor-targeting drugs.
AB - Class F receptors are considered valuable therapeutic targets due to their role in human disease, but structural changes accompanying receptor activation remain unexplored. Employing population and cancer genomics data, structural analyses, molecular dynamics simulations, resonance energy transfer-based approaches and mutagenesis, we identify a conserved basic amino acid in TM6 in Class F receptors that acts as a molecular switch to mediate receptor activation. Across all tested Class F receptors (FZD 4,5,6,7, SMO), mutation of the molecular switch confers an increased potency of agonists by stabilizing an active conformation as assessed by engineered mini G proteins as conformational sensors. Disruption of the switch abrogates the functional interaction between FZDs and the phosphoprotein Dishevelled, supporting conformational selection as a prerequisite for functional selectivity. Our studies reveal the molecular basis of a common activation mechanism conserved in all Class F receptors, which facilitates assay development and future discovery of Class F receptor-targeting drugs.
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U2 - 10.1038/s41467-019-08630-2
DO - 10.1038/s41467-019-08630-2
M3 - Article
C2 - 30737406
AN - SCOPUS:85061253455
SN - 2041-1723
VL - 10
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 667
ER -
