Characterization of inverted membrane vesicles from the halophilic archaeon Haloferax volcanii

G. Ring, J. Eichler

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Membrane-related processes in archaea, the third and most-recently described domain of life, are in general only poorly understood. One obstacle to a functional understanding of archaeal membrane-associated activities corresponds to a lack of archaeal model membrane systems. In the following, characterization of inverted archaeal membrane vesicles, prepared from the halophilic archaeon Haloferax volcanii, is presented. The inverted topology of the vesicles was revealed by defining the orientation of membrane-bound enzymes that in intact cells normally face the cytoplasm or of other protein markers, known to face the exterior medium in intact cells. Electron microscopy, protease protection assays and lectin-binding experiments confirmed the sealed nature of the vesicles. Upon alkalinization of the external medium, the vesicles were able to generate ATP, reflecting the functional nature of the membrane preparation. The availability of preparative scale amounts of inverted archaeal membrane vesicles provides a platform for the study of various membrane-related phenomena in archaea.

Original languageEnglish
Pages (from-to)195-204
Number of pages10
JournalJournal of Membrane Biology
Volume183
Issue number3
DOIs
StatePublished - 1 Oct 2001

Keywords

  • Archaea
  • Haloferax volcanii
  • Halophiles
  • Inverted membrane vesicles
  • Membrane proteins
  • Membrane transport

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