Journal Article

Smoothened stimulation by membrane sterols drives Hedgehog pathway activity

Faust, Bryan, Beachy, Philip A., Dror, Ron O., Roberts, Kelsey J., Zhang, Yunxiao, Manglik, Aashish, Ha, Betty, Deshpande, Ishan, Latorraca, Naomi R., Myers, Benjamin R., Liang, Jiahao, Hedeen, Danielle

2019

Overview

Hedgehog signalling is fundamental to embryonic development and postnatal tissue regeneration 1 . Aberrant postnatal Hedgehog signalling leads to several malignancies, including basal cell carcinoma and paediatric medulloblastoma 2 . Hedgehog proteins bind to and inhibit the transmembrane cholesterol transporter Patched-1 (PTCH1), which permits activation of the seven-transmembrane transducer Smoothened (SMO) via a mechanism that is poorly understood. Here we report the crystal structure of active mouse SMO bound to both the agonist SAG21k and to an intracellular binding nanobody that stabilizes a physiologically relevant active state. Analogous to other G protein-coupled receptors, the activation of SMO is associated with subtle motions in the extracellular domain, and larger intracellular changes. In contrast to recent models 3 – 5 , a cholesterol molecule that is critical for SMO activation is bound deep within the seven-transmembrane pocket. We propose that the inactivation of PTCH1 by Hedgehog allows a transmembrane sterol to access this seven-transmembrane site (potentially through a hydrophobic tunnel), which drives the activation of SMO. These results—combined with signalling studies and molecular dynamics simulations—delineate the structural basis for PTCH1–SMO regulation, and suggest a strategy for overcoming clinical resistance to SMO inhibitors. The crystal structure of active mouse SMO in complex with the SAG21k agonist and a stabilizing intracellular binding nanobody reveals the structural basis of SMO regulation by PTCH1.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/136 / 631/45/287/1197 / 631/45/612/194 / 631/535/1266 / 631/80/86/2370 / Activation / Animals / Basal cell carcinoma / BASIC BIOLOGICAL SCIENCES / Binding Sites / Biosensing Techniques / Cancer / Catalytic Domain - drug effects / Cell Membrane - chemistry / Cell Membrane - metabolism / Cholesterol / Cholesterol - chemistry / Cholesterol - metabolism / Cholesterol - pharmacology / Computer simulation / Crystal structure / Deactivation / Development and progression / Developmental biology / Drug resistance / Embryogenesis / Embryonic growth stage / G protein-coupled receptors / Hedgehog protein / Hedgehog proteins / Hedgehog Proteins - agonists / Hedgehog Proteins - metabolism / Humanities and Social Sciences / Hydrophobicity / Inactivation / Intracellular / Letter / Ligands / Medulloblastoma / Mice / Models, Molecular / Molecular dynamics / Molecular Dynamics Simulation / Morphogen signalling / multidisciplinary / Mutation / Nanobodies / Patched protein / Patched-1 Receptor - antagonists & inhibitors / Patched-1 Receptor - metabolism / Physiological aspects / Physiology / Plasma / Protein Conformation / Protein Stability / Proteins / Receptors / Regeneration / Science / Science (multidisciplinary) / Signal transduction / Signal Transduction - drug effects / Signaling / Single-Chain Antibodies - immunology / Skin cancer / Smoothened Receptor - agonists / Smoothened Receptor - antagonists & inhibitors / Smoothened Receptor - chemistry / Smoothened Receptor - metabolism / Software / Sterols / Sterols - chemistry / Sterols - metabolism / Sterols - pharmacology / Tissue engineering / Tumors / X-ray crystallography / Xenopus Proteins - chemistry