Fusion of native or reconstituted membranes to liposomes, optimized for single channel recording

A. M. Correa, W. S. Agnew

Research output: Contribution to journalArticle

Abstract

We here describe a protocol for fusing vesicles into large structures suitable for patch clamp recording. The method may be used with native membrane vesicles or with liposomes containing reconstituted/purified ion channels. The resulting unilamellar membranes exhibit high channel surface abundance, yielding multiple channels in the average excised patch. The procedure has been used to record voltage-sensitive Na channels from three native membrane preparations (eel electroplax, rat skeletal muscle, squid optic nerve), and from reconstituted protein purified from eel electroplax. Channels treated with batrachotoxin (BTX) displayed characteristic activation voltage dependence, conductances, selectivity, and sensitivity to saxitoxin (STX).

Original languageEnglish (US)
Pages (from-to)569-575
Number of pages7
JournalBiophysical journal
Volume54
Issue number3
DOIs
StatePublished - Jan 1 1988
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics

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