Experimental determination of control by the H+-ATPase in Escherichia coli

Peter Ruhdal Jensen, Ole Michelsen, H. V. Westerhoff

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Strains carrying deletions in the atp genes, encoding the H+-ATPase, were unable to grow on nonfermentable substrates such as succinate, whereas with glucose as the substrate the growth rate of an atp deletion mutant was surprisingly high (some 75-80% of wild-type growth rate). The rate of glucose and oxygen consumption of these mutants was increased compared to the wild-type rates. In order to analyze the importance of the H+-ATPase at its physiological level, the cellular concentration of H+-ATPase was modulated around the wild-type level, using genetically manipulated strains. The control coefficient by the H+-ATPase with respect to growth rate and catabolic fluxes was measured. Control on growth rate was absent at the wildtype concentration of H+-ATPase, independent of whether the substrate for growth was glucose or succinate. Control by the H+-ATPase on the catabolic fluxes, including respiration, was negative at the wild-type H+-ATPase level. Moreover, the turnover number of the individual H+-ATPase enzymes increased as the H+-ATPase concentration was lowered. The negative control by the H+-ATPase on catabolism may thus be involved in a homeostatic control of ATP synthesis and, to some extent, explain the zero control by the H+-ATPase on E. coli growth rate.
    Original languageEnglish
    JournalJournal of Bioenergetics and Biomembranes
    Volume27
    Issue number6
    Pages (from-to)543-554
    Number of pages12
    ISSN0145-479X
    DOIs
    Publication statusPublished - 1995

    Keywords

    • Escherichia coli
    • Gene Deletion
    • Genes, Bacterial
    • Glucose
    • Kinetics
    • Operon
    • Oxygen Consumption
    • Proton-Translocating ATPases
    • Succinates
    • EC 3.6.3.14 Proton-Translocating ATPases
    • IY9XDZ35W2 Glucose
    • BIOPHYSICS
    • CELL
    • PROTON-TRANSLOCATING ATPASE
    • NUCLEOTIDE-SEQUENCE
    • OXIDATIVE-PHOSPHORYLATION
    • ADENOSINE-TRIPHOSPHATASE
    • SYNTHASE
    • OPERON
    • GENES
    • PROMOTERS
    • SUBUNIT
    • MUTATIONS
    • oxidative phosphorylation
    • respiration
    • ATP
    • fluxes
    • Metabolic Control Analysis
    • Oxidative phosphorylation
    • ATP GENE
    • BACTERIAL GROWTH RATE
    • CATABOLIC FLUX
    • GENETICS
    • MUTATION
    • OXIDATIVE PHOSPHORYLATION
    • OXYGEN CONSUMPTION
    • RESPIRATION
    • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Escherichia coli
    • ATP 56-65-5Q, 42530-29-0Q, 94587-45-8Q, 111839-44-2Q
    • ATPASE 9000-83-3
    • OXYGEN 7782-44-7
    • 02506, Cytology - Animal
    • 10012, Biochemistry - Gases
    • 10060, Biochemistry studies - General
    • 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
    • 10510, Biophysics - Bioenergetics: electron transport and oxidative phosphorylation
    • 10808, Enzymes - Physiological studies
    • 13003, Metabolism - Energy and respiratory metabolism
    • 16004, Respiratory system - Physiology and biochemistry
    • 31000, Physiology and biochemistry of bacteria
    • 31500, Genetics of bacteria and viruses
    • 36002, Medical and clinical microbiology - Bacteriology
    • Biochemistry and Molecular Biophysics
    • Bioenergetics
    • Cell Biology
    • Enzymology
    • Genetics
    • Infection
    • Metabolism
    • Physiology

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