title: Aerobic Growth of Escherichia coli Is Reduced, and ATP Synthesis Is Selectively Inhibited when Five C-terminal Residues Are Deleted from the ϵ Subunit of ATP Synthase.
abstract: F-type ATP synthases are rotary nanomotor enzymes involved in cellular energy metabolism in eukaryotes and eubacteria. The ATP synthase from Gram-positive and -negative model bacteria can be autoinhibited by the C-terminal domain of its ϵ subunit (ϵCTD), but the importance of ϵ inhibition in vivo is unclear. Functional rotation is thought to be blocked by insertion of the latter half of the ϵCTD into the central cavity of the catalytic complex (F1). In the inhibited state of the Escherichia coli enzyme, the final segment of ϵCTD is deeply buried but has few specific interactions with other subunits. This region of the ϵCTD is variable or absent in other bacteria that exhibit strong ϵ-inhibition in vitro. Here, genetically deleting the last five residues of the ϵCTD (ϵΔ5) caused a greater defect in respiratory growth than did the complete absence of the ϵCTD. Isolated membranes with ϵΔ5 generated proton-motive force by respiration as effectively as with wild-type ϵ but showed a nearly 3-fold decrease in ATP synthesis rate. In contrast, the ϵΔ5 truncation did not change the intrinsic rate of ATP hydrolysis with membranes. Further, the ϵΔ5 subunit retained high affinity for isolated F1 but reduced the maximal inhibition of F1-ATPase by ϵ from >90% to ∼20%. The results suggest that the ϵCTD has distinct regulatory interactions with F1 when rotary catalysis operates in opposite directions for the hydrolysis or synthesis of ATP.
authors.affiliation: Department of Biochemistry and Molecular Biology, State University of New York, Upstate Medical University, Syracuse, New York 13210.
authors.forename: Naman B
authors.initials: NB
authors.lastname: Shah
authors.affiliation: Department of Biochemistry and Molecular Biology, State University of New York, Upstate Medical University, Syracuse, New York 13210. Electronic address: duncant@upstate.edu.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Duncan
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D055162:Biocatalysis; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018892:Proton-Motive Force; D011522:Protons; D017384:Sequence Deletion; D013816:Thermodynamics; D000097795:ATPase Inhibitory Protein
chemical_list: D000255:Adenosine Triphosphate; D029968:Escherichia coli Proteins; D011506:Proteins; D011522:Protons; D000097795:ATPase Inhibitory Protein
keywords: ATP synthase; Escherichia coli (E. coli); antibiotics; bioenergetics; enzyme inhibitor; epsilon subunit; mutagenesis; oxidative phosphorylation; proton-translocating ATPases