Structural basis of frizzled 7 activation and allosteric regulation

Frizzleds (ten paralogs: FZD 1-10 ) belong to the class F of G protein-coupled receptors (GPCRs), which remains poorly understood despite its crucial role in multiple key biological functions including embryonic development, stem cell regulation, and homeostasis in the adult. FZD 7 , one of the most studied members of the family, is more specifically involved in the migration of mesendoderm cells during the development and renewal of intestinal stem cells in adults. Moreover, FZD 7 has been highlighted for its involvement in tumor development predominantly in the gastrointestinal tract. This study reports the structure of inactive FZD 7 , without any stabilizing mutations, determined by cryo-electron microscopy (cryo-EM) at 1.9 Å resolution. We characterize a fluctuating water pocket in the core of the receptor important for FZD 7 dynamics. Molecular dynamics simulations are used to investigate the temporal distribution of those water molecules and their importance for potential conformational changes in FZD 7 . Moreover, we identify lipids interacting with the receptor core and a conserved cholesterol-binding site, which displays a key role in FZD 7 association with a transducer protein, Disheveled (DVL), and initiation of downstream signaling and signalosome formation. FZD 7 is a class F GPCR involved in intestinal epithelium homeostasis. Using cryo-EM, the authors determine the structure of inactive FZD 7 and compare it with the G-protein-bound form. They refine the FZD activation mechanisms and identify a water pocket and an allosteric cholesterol binding site.