Substitution of a single amino acid switches the tentoxin-resistant thermophilic F1-ATPase into a tentoxin-sensitive enzyme

J Biol Chem. 2002 Jun 7;277(23):20117-9. doi: 10.1074/jbc.C200168200. Epub 2002 Apr 9.

Abstract

In contrast to the homologous bacterial and mitochondrial enzymes the chloroplast F(1)-ATPase (CF(1)) is strongly affected by the phytopathogenic inhibitor tentoxin. Based on structural information obtained from crystals of a CF(1)-tentoxin co-complex (Groth, G. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 3464-3468) we have replaced residues betaSer(66) and alphaArg(132) in the alpha(3)beta(3)gamma subcomplex of the thermophilic F(1)-ATPase from Bacillus PS3 by the corresponding residues of the chloroplast ATPase to confer tentoxin sensitivity to the thermophilic enzyme. The mutation alphaArg(132) --> Pro, proposed to relieve steric constraints on tentoxin binding, did not have any significant effect. However, mutation betaSer(66) --> Ala, predicted to provide a crucial hydrogen bond with the inhibitor, resulted in tentoxin inhibition of ATP hydrolysis comparable with the situation found with the chloroplast enzyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Amino Acid Substitution*
  • Base Sequence
  • DNA Primers
  • Hydrolysis
  • Models, Molecular
  • Peptides, Cyclic / pharmacology*
  • Protein Conformation
  • Proton-Translocating ATPases / chemistry
  • Proton-Translocating ATPases / drug effects*
  • Proton-Translocating ATPases / metabolism

Substances

  • DNA Primers
  • Peptides, Cyclic
  • Adenosine Triphosphate
  • Proton-Translocating ATPases
  • tentoxin