Three-dimensional structure of A1A0 ATP synthase from the hyperthermophilic archaeon Pyrococcus furiosus by electron microscopy

J Biol Chem. 2009 Apr 10;284(15):10110-9. doi: 10.1074/jbc.M808498200. Epub 2009 Feb 8.

Abstract

The archaeal ATP synthase is a multisubunit complex that consists of a catalytic A(1) part and a transmembrane, ion translocation domain A(0). The A(1)A(0) complex from the hyperthermophile Pyrococcus furiosus was isolated. Mass analysis of the complex by laser-induced liquid bead ion desorption (LILBID) indicated a size of 730 +/- 10 kDa. A three-dimensional map was generated by electron microscopy from negatively stained images. The map at a resolution of 2.3 nm shows the A(1) and A(0) domain, connected by a central stalk and two peripheral stalks, one of which is connected to A(0), and both connected to A(1) via prominent knobs. X-ray structures of subunits from related proteins were fitted to the map. On the basis of the fitting and the LILBID analysis, a structural model is presented with the stoichiometry A(3)B(3)CDE(2)FH(2)ac(10).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Amino Acid Sequence
  • Archaea / enzymology
  • Crystallography, X-Ray / methods
  • Ions
  • Mass Spectrometry / methods
  • Microscopy, Electron / methods
  • Molecular Conformation
  • Molecular Sequence Data
  • Protein Structure, Secondary
  • Proton-Translocating ATPases / chemistry*
  • Pyrococcus furiosus / enzymology*
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Trypsin / chemistry

Substances

  • Ions
  • Adenosine Triphosphate
  • Trypsin
  • Proton-Translocating ATPases