The F1F0-ATPase of Escherichia coli. The substitution of alanine by tyrosine at position 25 in the c-subunit affects function but not assembly

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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The F1F0-ATPase of Escherichia coli. The substitution of glycine by valine at position 29 in the c-subunit affects function but not assembly

Biochimica et Biophysica Acta (BBA) - Biomembranes ◽

10.1016/0005-2736(89)90475-6 ◽

1989 ◽

Vol 986 (2) ◽

pp. 257-262 ◽

Cited By ~ 3

Author(s):

Anthony L. Fimmel ◽

Ursula Norris

Keyword(s):

Escherichia Coli ◽

F1f0 Atpase ◽

C Subunit

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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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Effect of the delta subunit on assembly and proton permeability of the F0 proton channel of Escherichia coli F1F0 ATPase.

Journal of Bacteriology ◽

10.1128/jb.173.1.407-411.1991 ◽

1991 ◽

Vol 173 (1) ◽

pp. 407-411 ◽

Cited By ~ 10

Author(s):

E Angov ◽

T C Ng ◽

W S Brusilow

Keyword(s):

Escherichia Coli ◽

Proton Channel ◽

Proton Permeability ◽

F1f0 Atpase ◽

Delta Subunit

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F1F0-ATPase from Escherichia coli with mutant F0 subunits. Partial purification and immunoprecipitation of F1F0 complexes.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)47569-2 ◽

1987 ◽

Vol 262 (17) ◽

pp. 8340-8346 ◽

Cited By ~ 3

Author(s):

S B Vik ◽

R D Simoni

Keyword(s):

Escherichia Coli ◽

Partial Purification ◽

F1f0 Atpase

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Mutations in the uncE gene affecting assembly of the c-subunit of the adenosine triphosphatase of Escherichia coli

Biochemical Journal ◽

10.1042/bj2110717 ◽

1983 ◽

Vol 211 (3) ◽

pp. 717-726 ◽

Cited By ~ 37

Author(s):

D A Jans ◽

A L Fimmel ◽

L Langman ◽

L B James ◽

J A Downie ◽

...

Keyword(s):

Escherichia Coli ◽

Amino Acid ◽

Cell Membrane ◽

Dna Sequences ◽

Adenosine Triphosphatase ◽

Gene Dosage ◽

Base Change ◽

Amino Acid Substitutions ◽

Helical Hairpin ◽

C Subunit

The amino acid substitutions in the mutant c-subunits of Escherichia coli F1F0-ATPase coded for by the uncE429, uncE408 and uncE463 alleles affect the incorporation of these proteins into the cell membrane. The DNA sequence of the uncE429 allele differed from normal in that a G leads to A base change occurred at nucleotide 68 of the uncE gene, resulting in glycine being replaced by aspartic acid at position 23 in the c-subunit. The uncE408 and uncE463 mutant DNA sequences were identical and differed from normal in that a C leads to T base change occurred at nucleotide 91 of the uncE gene, resulting in leucine being replaced by phenylalanine at position 31 in the c-subunit. An increased gene dosage of the uncE408 or uncE463 alleles resulted in the incorporation into the membranes of the mutant c-subunits. The results are discussed in terms of the ‘Helical Hairpin Hypothesis’ of Engelman & Steitz [(1981) Cell 23,411-422].

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