A mechanism of proton translocation by F1F0 ATP synthases suggested by double mutants of the a subunit.
Three amino acid residues in the a subunit of the Escherichia coli F1F0 ATP synthase are essential for proton translocation: Arg210, Glu219, and His245. In this study, the essential glutamic acid has been relocated to position 252 with retention of function. It had been known that Gln252 can be replaced by Glu without significant effect. To test whether Q252E would function in the absence of Glu219, a "site-directed second-site suppressor" experiment was designed. Saturation mutagenesis was applied to residue Glu219, and 14 different amino acid substitutions were isolated, five of which permitted growth on succinate minimal medium at 37 degrees C: Asp, Lys, Gly, Ala, and Ser. These results indicate that Q252E can provide the essential carboxyl group normally provided by Glu219, but that strict requirements are placed on the residue at position 219. We interpret these results to mean that the Q252E must occupy, at least partially, the normal position of Glu219. We present a novel mechanism of proton translocation by F1F0 ATP synthases that includes a rotating oligomer of c subunits, in which the Asp61 of two c subunits simultaneously interact with Glu219 and Arg210 of the a subunit. This mechanism can be adapted for both mitochondrial and sodium-driven bacterial ATP synthases.
Vik
S B
SB
Department of Biological Sciences, Southern Methodist University, Dallas, Texas 75275.
Antonio
B J
BJ
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D001483:Base Sequence; D002417:Cattle; D016384:Consensus Sequence; D004926:Escherichia coli / Q000201:enzymology*; D018698:Glutamic Acid; D005973:Glutamine; D006639:Histidine; D007700:Kinetics; D046911:Macromolecular Substances; D008961:Models, Structural; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D005973:Glutamine; D018698:Glutamic Acid; D006639:Histidine; D001120:Arginine; D006180:Proton-Translocating ATPases