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Related Experiment Videos

Decrease of NADH in HeLa cells in the presence of transferrin or ferricyanide.

P Navas, I L Sun, D J Morré

    Biochemical and Biophysical Research Communications
    |February 26, 1986
    PubMed
    Summary

    HeLa cells utilize NADH as an internal electron donor for transplasma membrane reductase, which reduces external oxidants like ferricyanide. This process, stimulated by insulin, appears sufficient to promote cell growth.

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    Area of Science:

    • Cellular Metabolism
    • Biochemistry
    • Molecular Biology

    Background:

    • Cellular respiration involves electron transport chains.
    • Pyridine nucleotides (NAD+, NADH, NADP+, NADPH) are crucial redox cofactors.
    • Transmembrane electron transport is a less understood cellular process.

    Purpose of the Study:

    • To investigate the role of NADH in transplasma membrane electron transport.
    • To determine if external oxidants can be reduced by intracellular NADH in HeLa cells.
    • To explore the effect of insulin on this process.

    Main Methods:

    • Incubation of HeLa cells with specific oxidants (diferric transferrin, ferricyanide) and controls (apotransferrin, ferrocyanide).
    • Measurement of intracellular NAD+ and NADH concentrations.

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  • Assessment of the NADP/NADPH ratio and total pyridine nucleotide pool.

    • Main Results:

    • Incubation with oxidants caused a stoichiometric decrease in NADH and increase in NAD+.
    • Insulin significantly stimulated this NADH oxidation.
    • The NADP/NADPH ratio and total pyridine nucleotide pool remained unchanged.
    • Control incubations without effective oxidants did not alter pyridine nucleotide levels.

    Conclusions:

    • NADH serves as the internal electron donor for the transplasma membrane reductase.
    • The transmembrane electron transport utilizing ferricyanide or diferric transferrin as acceptors is sufficient to stimulate HeLa cell growth.