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Phosphorylative fumarate reduction in Vibrio succinogenes: Stoichiometry of ATP synthesis

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Abstract

  1. 1.

    Cells of Vibrio succinogenes, treated with EDTA at pH 8, catalyze the phosphorylation of their endogenous ADP and AMP as a function of the electron transport from formate to fumarate. The P/fumarate ratio obtained from the initial velocity of the phosphorylation on initiation of the electron transport and from the activity of fumarate reduction in the steady state was 0.90. The phosphorylation was prevented by 10μmol/g protein carbonylcyanide-3-chlorophenylhydrazone.

  2. 2.

    The esterification of external phosphate in the presence of ADP, hexokinase and glucose is catalysed by a membrane preparation of V. succinogenes in the steady state of fumarate reduction by H2. The phosphorylation was fully abolished by either 5μmol/g protein carbonylcyanide-4-trifluoromethoxyphenylhydrazone or 30μmol/g protein carbonylcyanide-3-chlorphenylhydrazone. Phosphorylation was blocked also by dicyclohexylcarbodiimide, an inhibitor of the Mg2+-dependent membrane bound ATP synthase, and by low concentrations of the inhibitors of electron transport 2-(n-nonyl)-4-hydroxyquinoline-N-oxide or 4-chloromercuriphenylsulfonate.

  3. 3.

    The P/fumarate ratios, measured with the membrane preparation, were found to increase with progressive inhibition of the electron transport from hydrogen to fumarate by means of 4-chloromercuriphenylsulfonate. The extrapolated ratio at vanishing electron transport activity was 0.47.

  4. 4.

    About 50% of the membrane preparation was found to consist of inverted vesicles with the hydrogenase and formate dehydrogenase oriented to the inside. The residual part is considered as being incapable of performing energy transduction. The extrapolated P/fumarate ratio valid for the inverted vesicles was 0.94.

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Abbreviations

EDTA:

ethylene diamine tetraacetate

MES:

2-(N-Morpholino)ethanesulfonate

TRIS:

Tris(hydroxymethyl)aminomethane

HEPES:

N-2-Hydroxyethylpiperazine-N′-2-ethanesulfonate

CCCP:

Carbonylcyanide-3-chlorophenylhydrazone

FCCP:

Carbonylcyanide-4-trifluoromethoxyphenylhydrazone

p-CMS:

4-Chloromercuriphenylsulfonate

NQNO:

2-(n-Nonyl)-4-hydroxyquinoline-N-oxide

DCCD:

N,N′-Dicyclohexylcarbodiimide

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Authors and Affiliations

  1. Institut für Physiologische Chemie der Universität München, Goethestr. 33, D-8000, München 2, Germany

    Edith Winkler

  2. Fachbereich Biologie/Mikrobiologie, Philipps-Universität Marburg, Lahnberge, D-3550, Marburg, Germany

    Achim Kröger

Authors
  1. Achim Kröger
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  2. Edith Winkler
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Kröger, A., Winkler, E. Phosphorylative fumarate reduction in Vibrio succinogenes: Stoichiometry of ATP synthesis. Arch. Microbiol. 129, 100–104 (1981). https://doi.org/10.1007/BF00417188

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  • DOI: https://doi.org/10.1007/BF00417188

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