The structure and nucleotide occupancy of bovine mitochondrial F-1-ATPase are not influenced by crystallisation at high concentrations of nucleotide

Analysis of tryptophan mutants of F-1-ATPase from Escherichia coli [Lobau e t al, (1997) FEES Lett, 404, 15-18] suggested that nucleotide concentration s used to grow crystals for the determination of the structure of bovine F- 1-ATPase [Abrahams et al, (1994) Nature 370, 621-628] would be sufficient t o occupy only two catalytic sites, and that higher concentrations of nucleo tide would result in all three sites being occupied. We have determined the structure of bovine F-1-ATPase at 2.9 Angstrom resolution with crystals gr own in the presence of 5 mM AMPPNP and 5 muM ADP, Similar to previous struc tures of bovine F-1-ATPase determined with crystals grown in the presence o f lower nucleotide concentrations, only two beta -subunits have bound nucle otide and the third subunit remains empty. (C) 2001 Published by Elsevier S cience B,V, on behalf of the Federation of European Biochemical Societies.