Studies of SMOOTHENED Activation in Cell-Free and Reconstituted Systems

Methods Mol Biol. 2022:2374:161-174. doi: 10.1007/978-1-0716-1701-4_14.

Abstract

Much of our current understanding of Hedgehog signal transduction derives from studies involving intact cells and organisms. Here we describe the use of cell-free and reconstituted systems to study a key step in Hedgehog signal transduction: the activation of SMOOTHENED by membrane lipids. These methods can be adapted to study other steps in Hedgehog signal transduction, particularly those that occur at the membrane.

Keywords: Cholesterol; G protein; GPCR; Hedgehog; Lipid; Membrane; Nanodisc; PATCHED; Reconstitution; SMOOTHENED.

Publication types

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

MeSH terms

  • Cell-Free System
  • Hedgehog Proteins
  • Receptors, Cell Surface / metabolism*
  • Receptors, G-Protein-Coupled
  • Signal Transduction
  • Smoothened Receptor

Substances

  • Hedgehog Proteins
  • Receptors, Cell Surface
  • Receptors, G-Protein-Coupled
  • Smoothened Receptor