Resolution of the V1 ATPase from Manduca sexta into subcomplexes and visualization of an ATPase-active A3B3EG complex by electron microscopy

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Vincenzo F. Rizzo - , Saarland University (Author)
  • Ünal Coskun - , Saarland University (Author)
  • Michael Radermacher - , University of Vermont (Author)
  • Teresa Ruiz - , University of Vermont (Author)
  • Andrea Armbrüster - , Saarland University (Author)
  • Gerhard Grüber - , Saarland University (Author)

Abstract

The effect of the ATPase activity of Manduca sexta V1 ATPase by the amphipathic detergent lauryldimethylamine oxide (LDAO) and the relationship of these activities to the subunit composition of V1 were studied. The V1 was highly activated in the presence of 0.04-0.06% LDAO combined with release of the subunits H, C, and F from the enzyme. Increase of LDAO concentration to 0.1-0.2% caused the characterized subcomplexes A3B3HEGF and A3B3EG with a remaining ATPase activity of 52 and 65%, respectively. The hydrolytic-active A3B3EG subcomplex has been visualized by electron microscopy showing six major masses of density in a pseudo-hexagonal arrangement surrounding a seventh mass. The compositions of the various subcomplexes and fragments of V1 provide an organization of the subunits in the enzyme in the framework of the known three-dimensional reconstruction of the V1 ATPase from M. sexta (Radermacher, M., Ruiz, T., Wieczorek, H., and Grüber, G. (2001) J. Struct. Biol. 135, 26-37).

Details

Original languageEnglish
Pages (from-to)270-275
Number of pages6
JournalJournal of Biological Chemistry
Volume278
Issue number1
Publication statusPublished - 3 Jan 2003
Peer-reviewedYes
Externally publishedYes

External IDs

WOS 000180255700036
Scopus 0037414640
PubMed 12414800

Keywords

Library keywords