Incorporation of reconstituted acetylcholine receptors from Torpedo into the Xenopus oocyte membrane

Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8468-72. doi: 10.1073/pnas.92.18.8468.

Abstract

Xenopus oocytes are a valuable aid for studying the molecular structure and function of ionic channels and neurotransmitter receptors. Their use has recently been extended by the demonstration that oocytes can incorporate foreign membranes carrying preassembled receptors and channels. Here we show that when reconstituted in an artificial lipid matrix and injected into Xenopus oocytes, purified nicotinic acetylcholine receptors are efficiently inserted into the plasma membrane, where they form "clusters" of receptors that retain their native properties. This constitutes an innovative approach that, besides allowing the analyses of membrane fusion processes, is also a powerful technique for studying the characteristics and regulation of many membrane proteins (with their native stoichiometry and configuration) upon reinsertion into the membrane of a very convenient host cell system.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Cell Membrane / physiology
  • Membrane Potentials / physiology
  • Microinjections
  • Oocytes / metabolism
  • Oocytes / physiology
  • Receptors, Cholinergic / genetics
  • Receptors, Cholinergic / metabolism
  • Receptors, Cholinergic / physiology*
  • Torpedo
  • Xenopus laevis

Substances

  • Receptors, Cholinergic