Mechanical Unfolding and Refolding of Single Membrane Proteins by Atomic Force Microscopy

Methods Mol Biol. 2020:2127:359-372. doi: 10.1007/978-1-0716-0373-4_23.

Abstract

Atomic force microscopy (AFM)-based single-molecule force spectroscopy allows direct physical manipulation of single membrane proteins under near-physiological conditions. It can be applied to study mechanical properties and molecular interactions as well as unfolding and folding pathways of membrane proteins. Here, we describe the basic procedure to study membrane proteins by single-molecule force spectroscopy and discuss general requirements of the experimental setup as well as common pitfalls typically encountered when working with membrane proteins in AFM.

Keywords: Atomic force microscopy; Mechanical unfolding; Membrane protein folding; Single-molecule force spectroscopy; Supported lipid bilayer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriorhodopsins / chemistry
  • Haloarcula / chemistry
  • Haloarcula / metabolism
  • Humans
  • Mechanical Phenomena
  • Membrane Proteins / chemistry*
  • Microscopy, Atomic Force / methods*
  • Protein Refolding*
  • Protein Unfolding*
  • Proteolipids / chemistry
  • Single Molecule Imaging / methods*
  • Stress, Mechanical

Substances

  • Membrane Proteins
  • Proteolipids
  • proteoliposomes
  • Bacteriorhodopsins