Complexation of integral membrane proteins by phosphorylcholine-based amphipols

Biochim Biophys Acta. 2007 Nov;1768(11):2737-47. doi: 10.1016/j.bbamem.2007.07.007. Epub 2007 Jul 24.

Abstract

Amphiphilic macromolecules, known as amphipols, have emerged as promising candidates to replace conventional detergents for handling integral membrane proteins in water due to the enhanced stability of protein/amphipol complexes as compared to protein/detergent complexes. The limited portfolio of amphipols currently available prompted us to develop amphipols bearing phosphorylcholine-based units (PC). Unlike carboxylated polymers, PC-amphipols remain soluble in aqueous media under conditions of low pH, high salt concentration, or in the presence of divalent ions. The solubilizing properties of four PC-amphipols were assessed in the case of two membrane proteins, cytochrome b(6)f and bacteriorhodopsin. The protein/PC-amphipol complexes had a low dispersity in size, as determined by rate zonal ultracentrifugation. Short PC-amphipols (<M> approximately 22 kDa) of low dispersity in length, containing approximately 30 mol% octyl side groups, approximately 35 mol% PC-groups, and approximately 35 mol% isopropyl side groups, appeared best suited to form stable complexes, preserving the native state of BR over periods of several days. BR/PC-amphipol complexes remained soluble in aqueous media at pH> or =5, as well as in the presence of 1 M NaCl or 12 mM calcium ions. Results from isothermal titration calorimetry indicated that the energetics of the conversion of BR/detergent complexes into BR/amphipol complexes are similar for PC-amphipols and carboxylated amphiphols.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calorimetry
  • Detergents / chemistry*
  • Hydrogen-Ion Concentration
  • Membrane Proteins / chemistry*
  • Phosphorylcholine / chemistry*
  • Polymers / chemistry*
  • Propylamines / chemistry

Substances

  • Detergents
  • Membrane Proteins
  • Polymers
  • Propylamines
  • amphipol A8-35
  • Phosphorylcholine