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

Biochemistry ◽  
1990 ◽  
Vol 29 (45) ◽  
pp. 10387-10393 ◽  
Author(s):  
Robert Aggeler ◽  
Janet Mendel-Hartvig ◽  
Roderick A. Capaldi
Keyword(s):  
Escherichia Coli ◽  
Monoclonal Antibody ◽  
Atpase Activity ◽  
F1 Atpase

scholarly journals Oxidative phosphorylation by mutant Escherichia coli membranes with impaired proton permeability

1983 ◽  
Vol 216 (1) ◽  
pp. 143-150 ◽  
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.

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

1983 ◽  
Vol 213 (2) ◽  
pp. 451-458 ◽  
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.

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

1997 ◽  
Vol 83 (2) ◽  
pp. 132-138 ◽  
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

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

1983 ◽  
Vol 210 (2) ◽  
pp. 395-403 ◽  
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.

scholarly journals Reconstitution of F1 -ATPase activity from Escherichia coli subunits α, β and subunit γ tagged with six histidine residues at the C-terminus

FEBS Letters ◽  
1998 ◽  
Vol 427 (1) ◽  
pp. 64-68 ◽  
Author(s):  
Atsuko Ekuni ◽  
Hikaru Watanabe ◽  
Nozomi Kuroda ◽  
Ken Sawada ◽  
Hiroshi Murakami ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Atpase Activity ◽  
Histidine Residues ◽  
F1 Atpase ◽  
C Terminus

Binding of [14C]aurovertin D to Escherichia coli F1-ATPase and the isolated .beta. subunit. Correlation with inhibition of the ATPase activity

Biochemistry ◽  
1983 ◽  
Vol 22 (14) ◽  
pp. 3485-3492 ◽  
Author(s):  
Jean Paul Issartel ◽  
Gerard Klein ◽  
Michel Satre ◽  
Pierre V. Vignais
Keyword(s):  
Escherichia Coli ◽  
Atpase Activity ◽  
Beta Subunit ◽  
F1 Atpase

scholarly journals Properties of membranes from mutant strains of Escherichia coli in which the β-subunit of the adenosine triphosphatase is abnormal

1979 ◽  
Vol 180 (1) ◽  
pp. 111-118 ◽  
Author(s):  
A E Senior ◽  
D R Fayle ◽  
J A Downie ◽  
F Gibson ◽  
G B Cox
Keyword(s):  
Escherichia Coli ◽  
Atpase Activity ◽  
Mutant Allele ◽  
Adenosine Triphosphatase ◽  
Beta Subunit ◽  
Beta Subunits ◽  
F1 Atpase ◽  
Mutant Strains ◽  
Membrane Bound ◽  
Phenotypic Variations

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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