Crystal structure of a multi-domain human smoothened receptor in complex with a super stabilizing ligand

Nat Commun. 2017 May 17:8:15383. doi: 10.1038/ncomms15383.

Abstract

The Smoothened receptor (SMO) belongs to the Class Frizzled of the G protein-coupled receptor (GPCR) superfamily, constituting a key component of the Hedgehog signalling pathway. Here we report the crystal structure of the multi-domain human SMO, bound and stabilized by a designed tool ligand TC114, using an X-ray free-electron laser source at 2.9 Å. The structure reveals a precise arrangement of three distinct domains: a seven-transmembrane helices domain (TMD), a hinge domain (HD) and an intact extracellular cysteine-rich domain (CRD). This architecture enables allosteric interactions between the domains that are important for ligand recognition and receptor activation. By combining the structural data, molecular dynamics simulation, and hydrogen-deuterium-exchange analysis, we demonstrate that transmembrane helix VI, extracellular loop 3 and the HD play a central role in transmitting the signal employing a unique GPCR activation mechanism, distinct from other multi-domain GPCRs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Crystallography, X-Ray
  • Deuterium Exchange Measurement / methods
  • HEK293 Cells
  • Hedgehog Proteins / metabolism*
  • Humans
  • Ligands
  • Mass Spectrometry / methods
  • Molecular Dynamics Simulation
  • Protein Binding
  • Protein Domains*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Signal Transduction*
  • Smoothened Receptor / chemistry*
  • Smoothened Receptor / isolation & purification
  • Smoothened Receptor / metabolism

Substances

  • Hedgehog Proteins
  • Ligands
  • Recombinant Proteins
  • SMO protein, human
  • Smoothened Receptor