The proton pore of the F0F1-ATPase of Escherichia coli: Ser-206 is not required for proton translocation.
A series of experiments was carried out to investigate the role of some polar amino acids in the a-subunit of the ATP synthase of Escherichia coli. Site-directed mutagenesis resulted in the amino acid substitutions Ser-199----Ala, Ser-202----Ala, Ser-206----Ala, Arg-61----Gln or Asp-44----Asn. None of these amino acid substitutions affected the ability of the cells to carry out oxidative phosphorylation. It was concluded therefore that the effect of the substitution of leucine for Ser-206 reported previously (Cain, B.D. and Simoni, R.D. (1986) J. Biol. Chem. 261, 10043-10050) was due to the presence of the leucine rather than the absence of serine. Even though cells carrying the Asp-44----Asn substitution were able to carry out oxidative phosphorylation, membranes from such cells remained proton-impermeable after removal of the F1-ATPase. It appears likely that the proton pore of the F0 of the ATP synthase of E. coli consists of four amino acids, namely Arg-219, Glu-210 and His-245 of the a-subunit and Asp-61 of the c-subunit.
Howitt
S M
SM
Division of Biochemical Sciences, John Curtin School of Medical Research, Australian National University, Canberra.
Gibson
F
F
Cox
G B
GB
D000409:Alanine; D000595:Amino Acid Sequence; D001216:Asparagine; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005453:Fluorescence; D005973:Glutamine; D007473:Ion Channels / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D006180:Proton-Translocating ATPases / Q000378:metabolism; D011522:Protons; D011796:Quinacrine; D012694:Serine
D004269:DNA, Bacterial; D007473:Ion Channels; D011522:Protons; D005973:Glutamine; D012694:Serine; D004024:Dicyclohexylcarbodiimide; D001216:Asparagine; D004262:DNA Restriction Enzymes; D006180:Proton-Translocating ATPases; D011796:Quinacrine; D000409:Alanine