Accessible Mannitol-Based Amphiphiles (MNAs) for Membrane Protein Solubilisation and Stabilisation

Chemistry. 2016 May 17;22(21):7068-73. doi: 10.1002/chem.201600533. Epub 2016 Apr 13.

Abstract

Integral membrane proteins are amphipathic molecules crucial for all cellular life. The structural study of these macromolecules starts with protein extraction from the native membranes, followed by purification and crystallisation. Detergents are essential tools for these processes, but detergent-solubilised membrane proteins often denature and aggregate, resulting in loss of both structure and function. In this study, a novel class of agents, designated mannitol-based amphiphiles (MNAs), were prepared and characterised for their ability to solubilise and stabilise membrane proteins. Some of MNAs conferred enhanced stability to four membrane proteins including a G protein-coupled receptor (GPCR), the β2 adrenergic receptor (β2 AR), compared to both n-dodecyl-d-maltoside (DDM) and the other MNAs. These agents were also better than DDM for electron microscopy analysis of the β2 AR. The ease of preparation together with the enhanced membrane protein stabilisation efficacy demonstrates the value of these agents for future membrane protein research.

Keywords: amphiphile design; electron microscopy; membrane proteins; novel detergents; protein stabilization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiporters / chemistry
  • Antiporters / isolation & purification
  • Arabidopsis / chemistry
  • Arabidopsis Proteins / chemistry
  • Arabidopsis Proteins / isolation & purification
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / isolation & purification
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Light-Harvesting Protein Complexes / chemistry
  • Light-Harvesting Protein Complexes / isolation & purification
  • Mannitol / chemistry*
  • Membrane Proteins / chemistry*
  • Membrane Proteins / isolation & purification
  • Protein Stability
  • Receptors, Adrenergic, beta-2 / chemistry
  • Receptors, Adrenergic, beta-2 / isolation & purification
  • Rhodobacter capsulatus / chemistry
  • Solubility
  • Surface-Active Agents / chemistry*

Substances

  • Antiporters
  • Arabidopsis Proteins
  • BOR1 protein, Arabidopsis
  • Bacterial Proteins
  • Light-Harvesting Protein Complexes
  • Membrane Proteins
  • Receptors, Adrenergic, beta-2
  • Surface-Active Agents
  • light-harvesting complex 1, Rhodospirillum
  • Mannitol