Three-dimensional reconstruction of bovine brain V-ATPase by cryo-electron microscopy and single particle analysis

Marco Gregorini, Jin Wang, Xiao Song Xie, Ronald A. Milligan, Andreas Engel

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Bovine V-ATPase from brain clathrin-coated vesicles was investigated by cryo-electron microscopy and single particle analysis. Our studies revealed great flexibility of the central linker region connecting V1 and V0. As a consequence, the two sub-complexes were processed separately and the resulting volumes were merged computationally. We present the first three-dimensional (3D) map of a V-ATPase obtained from cryo-electron micrographs. The overall resolution was estimated 34 Å by Fourier shell correlation (0.5 cutoff). Our 3D reconstruction shows a large peripheral stalk and a smaller, isolated peripheral density, suggesting a second, less well-resolved peripheral connection. The 3D map reveals new features of the large peripheral stator and of the collar-like density attached to the membrane domain. Our analyses of the membrane domain indicate the presence of six proteolipid subunits. In addition, we could localize the V0 subunit a flanking the large peripheral stalk.

Original languageEnglish (US)
Pages (from-to)445-454
Number of pages10
JournalJournal of Structural Biology
Volume158
Issue number3
DOIs
StatePublished - Jun 1 2007

Keywords

  • 3D reconstruction
  • Bovine brain V-ATPase
  • Cryo-electron microscopy
  • Single particle analysis

ASJC Scopus subject areas

  • Structural Biology

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