Hybrid In Silico/In Vitro Approaches for the Identification of Functional Cholesterol-Binding Domains in Membrane Proteins

Methods Mol Biol. 2017;1583:7-19. doi: 10.1007/978-1-4939-6875-6_2.

Abstract

In eukaryotic cells, cholesterol is an important regulator of a broad range of membrane proteins, including receptors, transporters, and ion channels. Understanding how cholesterol interacts with membrane proteins is a difficult task because structural data of these proteins complexed with cholesterol are scarce. Here, we describe a dual approach based on in silico studies of protein-cholesterol interactions, combined with physico-chemical measurements of protein insertion into cholesterol-containing monolayers. Our algorithm is validated through careful analysis of the effect of key mutations within and outside the predicted cholesterol-binding site. Our method is illustrated by a complete analysis of cholesterol-binding to Alzheimer's β-amyloid peptide, a protein that penetrates the plasma membrane of brain cells through a cholesterol-dependent process.

Keywords: Alzheimer’s β-amyloid peptide; Cholesterol-binding motif; Langmuir monolayer; Molecular docking; Molecular dynamics simulations; Transmembrane domain.

MeSH terms

  • Algorithms*
  • Alzheimer Disease / genetics
  • Alzheimer Disease / metabolism
  • Amyloid beta-Peptides* / chemistry
  • Amyloid beta-Peptides* / genetics
  • Amyloid beta-Peptides* / metabolism
  • Animals
  • Cholesterol* / chemistry
  • Cholesterol* / genetics
  • Cholesterol* / metabolism
  • Computer Simulation*
  • Humans
  • Protein Domains
  • Sequence Analysis, Protein / methods*

Substances

  • Amyloid beta-Peptides
  • Cholesterol