Activation of Escherichia coli F1-ATPase by lauryldimethylamine oxide and ethylene glycol: relationship of ATPase activity to the interaction of the .epsilon. and .beta. subunits

Five uncoupled mutant strains of Escherichia coli carrying mutations in the uncD gene have been studied. In each of these mutant strains the beta-subunit of the F1 portion of the membrane-bound adenosine triphosphatase is abnormal. In one of the mutant strains (carrying the uncD12 allele) in F1-ATPase aggregate was formed which was purified and found to have low ATPase activity. ATPase activity was absent in the other four strains and the abnormal beta-subunits were tightly bound to the membranes. However, membranes from these strains exhibited various proton permeabilities as indicated by NADH-dependent atebrin-fluorescence quenching and bound different amounts of normal F1-ATPase. The amounts of reconstitution of energy-linked reactions after the addition of normal F1-ATPase also varied depending on the mutant allele. It is apparent that considerable phenotypic variations can occur between strains carrying mutations in the same unc gene.

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Oxidative phosphorylation by mutant Escherichia coli membranes with impaired proton permeability

Biochemical Journal ◽

10.1042/bj2160143 ◽

1983 ◽

Vol 216 (1) ◽

pp. 143-150 ◽

Cited By ~ 34

Author(s):

G B Cox ◽

D A Jans ◽

F Gibson ◽

L Langman ◽

A E Senior ◽

...

Keyword(s):

Escherichia Coli ◽

Oxidative Phosphorylation ◽

Atpase Activity ◽

Gene Dosage ◽

Control Strain ◽

Proton Permeability ◽

F1f0 Atpase ◽

F1 Atpase ◽

C Subunit

The effect on the function of the Escherichia coli F1F0-ATPase of the substitution of leucine-31 by phenylalanine in the c-subunit of the enzyme was examined. The assembly of the mutant c-subunit requires an increased gene dosage [Jans, Fimmel, Langman, James, Downie, Senior, Ash, Gibson & Cox (1983) Biochem. J. 211, 717-726], and this was achieved by incorporation of the uncE408 or uncE463 alleles on to F-plasmids or multicopy plasmids. Membranes from strains carrying either the uncE463 or uncE408 alleles on F-plasmids or multicopy plasmids were capable of carrying out oxidative phosphorylation. In particular, membranes from strain AN1928 (pAN162, uncE463) gave phosphorylation rates and P/O ratios equal to or greater than those obtained for the control strain AN1460 (pAN45, unc+). However, the mutant membranes, on removal of the F1-ATPase, appeared to be proton-impermeable. The ATPase activity of the mutant membranes was also resistant to the inhibitor dicyclohexylcarbodi-imide.

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Monoclonal antibody modification of the ATPase activity of Escherichia coli F1 ATPase

Biochemistry ◽

10.1021/bi00497a013 ◽

1990 ◽

Vol 29 (45) ◽

pp. 10387-10393 ◽

Cited By ~ 25

Author(s):

Robert Aggeler ◽

Janet Mendel-Hartvig ◽

Roderick A. Capaldi

Keyword(s):

Escherichia Coli ◽

Monoclonal Antibody ◽

Atpase Activity ◽

F1 Atpase

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Conformational interactions between alpha and beta subunits in the F1 ATPase of Escherichia coli as shown by chemical modification of uncA401 and uncD412 mutant enzymes

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1986.tb09954.x ◽

1986 ◽

Vol 160 (1) ◽

pp. 169-174 ◽

Cited By ~ 5

Author(s):

Helga STAN-LOTTER ◽

Philip D. BRAGG

Keyword(s):

Escherichia Coli ◽

Chemical Modification ◽

Beta Subunits ◽

F1 Atpase

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The F1F0-ATPase of Escherichia coli. Substitution of proline by leucine at position 64 in the c-subunit causes loss of oxidative phosphorylation

Biochemical Journal ◽

10.1042/bj2130451 ◽

1983 ◽

Vol 213 (2) ◽

pp. 451-458 ◽

Cited By ~ 33

Author(s):

A L Fimmel ◽

D A Jans ◽

L Langman ◽

L B James ◽

G R Ash ◽

...

Keyword(s):

Escherichia Coli ◽

Amino Acid ◽

Fluorescence Quenching ◽

Oxidative Phosphorylation ◽

Atpase Activity ◽

Base Change ◽

Helix Axis ◽

F1f0 Atpase ◽

F1 Atpase ◽

C Subunit

The uncE410 allele differs from the normal uncE gene in that C leads to T base changes occur at nucleotides 190 and 191, resulting in proline at position 64 in the c-subunit of the F1F0-ATPase being replaced by leucine. Two partial-revertant strains were isolated in which alanine-20 of the c-subunit was replaced by proline, owing to a G leads to C base change at nucleotide 58. These c-subunits, coded for by the uncE501 and uncE502 alleles, therefore contained two amino acid changes, namely proline-64 leads to leucine, and alanine-20 leads to proline. Membranes prepared from the partial-revertant strains lacked ATP-dependent atebrin-fluorescence-quenching activity but were able to carry out oxidative phosphorylation. The ATPase activity of the F1-ATPase was inhibited when bound to membranes from strains carrying the uncE410, uncE501 and uncE502 alleles. It is concluded that a bend in the helix axis in one of the arms of the c-subunit hairpin structure is required for integration of the c-subunit into a functional F1F0-ATPase.

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Isolation of α-subunits of factor F1 from submitochondrial particles and the reconstitution of active ATPase from isolated α-subunits and β-subunits bound to the mitochondrial membrane

Biochemical Journal ◽

10.1042/bj1920483 ◽

1980 ◽

Vol 192 (2) ◽

pp. 483-488 ◽

Cited By ~ 10

Author(s):

I A Kozlov ◽

Y M Milgrom ◽

I S Tsybovski

Keyword(s):

Atpase Activity ◽

Active Site ◽

Mitochondrial Membrane ◽

Iodoacetic Acid ◽

Reduced Form ◽

Beta Subunits ◽

Submitochondrial Particles ◽

F1 Atpase ◽

Alpha Subunits ◽

Α Subunits

The alpha-subunits of factor-F1 ATPase are removed by extraction of submitochondrial particles with 1.75 M-LiCl, with the consequent loss of ATPase activity. ATPase activity is reconstituted by incubation of LiCl-extracted particles with purified alpha-subunits, and the reconstituted ATPase activity is oligomycin-sensitive. Reconstitution is enhanced by maintenance of the alpha-subunits in reduced form by dithiothreitol or NaBH4 and by modification of the alpha-subunits by p-chloromercuribenzoate, iodoacetic acid or N-ethylmaleimide. Experiments with the mixed anhydride of ATP and mesitylene-carboxylic acid, which was previously shown to interact with the F1 active site, localized on the beta-subunits, indicate that the active site of ATPase is shielded by the alpha-subunits.

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Enhancement of glucose metabolism in a pyruvic acid-hyperproducing Escherichia coli mutant defective in F1-ATPase activity

Journal of Fermentation and Bioengineering ◽

10.1016/s0922-338x(97)83571-4 ◽

1997 ◽

Vol 83 (2) ◽

pp. 132-138 ◽

Cited By ~ 28

Author(s):

Atsushi Yokota ◽

Masaru Henmi ◽

Naohisa Takaoka ◽

Chiho Hayashi ◽

Yuji Takezawa ◽

...

Keyword(s):

Escherichia Coli ◽

Glucose Metabolism ◽

Atpase Activity ◽

Pyruvic Acid ◽

F1 Atpase ◽

Coli Mutant

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Oxidative phosphorylation in Escherichia coli. Characterization of mutant strains in which F1-ATPase contains abnormal β-subunits

Biochemical Journal ◽

10.1042/bj2100395 ◽

1983 ◽

Vol 210 (2) ◽

pp. 395-403 ◽

Cited By ~ 52

Author(s):

A E Senior ◽

L Langman ◽

G B Cox ◽

F Gibson

Keyword(s):

Escherichia Coli ◽

Ionic Strength ◽

Atpase Activity ◽

Atp Synthesis ◽

Beta Subunit ◽

F1 Atpase ◽

Dependent Atpase ◽

Mutant Strains ◽

Synthesis Activity ◽

Low Ionic Strength

To facilitate study of the role of the beta-subunit in the membrane-bound proton-translocating ATPase of Escherichia coli, we identified mutant strains from which an F1-ATPase containing abnormal beta-subunits can be purified. Seventeen strains of E. coli, characterized by genetic complementation tests as carrying mutations in the uncD gene (which codes for the beta-subunit), were studied. The majority of these strains (11) were judged to be not useful, as their membranes lacked ATPase activity, and were either proton-permeable as prepared or remained proton-impermeable after washing with buffer of low ionic strength. A further two strains were of a type not hitherto reported, in that their membranes had ATPase activity, were proton-impermeable as prepared, and were not rendered proton-permeable by washing in buffer of low ionic strength. Presumably in these two strains F1-ATPase is not released in soluble form by this procedure. F1-ATPase of normal molecular size were purified from strains AN1340 (uncD478), AN937 (uncD430), AN938 (uncD431) and AN1543 (uncD484). F1-ATPase from strain AN1340 (uncD478) had 15% of normal specific Mg-dependent ATPase activity and 22% of normal ATP-synthesis activity. The F1-ATPase preparations from strains AN937, AN938 and AN1543 had respectively 1.7%, 1.8% and 0.2% of normal specific Mg-dependent ATPase activity, and each of these preparations had very low ATP-synthesis activity. The yield of F1-ATPase from the four strains described was almost twice that obtained from a normal haploid strain. The kinetics of Ca-dependent ATPase activity were unusual in each of the four F1-ATPase preparations. It is likely that these four mutant uncD F1-ATPase preparations will prove valuable for further experimental study of the F1-ATPase catalytic mechanism.

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