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Related Experiment Videos

The lysosomal proton pump is electrogenic.

P Harikumar, J P Reeves

    The Journal of Biological Chemistry
    |September 10, 1983
    PubMed
    Summary

    Lysosomes generate an electrical potential across their membrane, driven by a proton pump. This electrogenic proton pump is crucial for lysosomal function and is modulated by ATP.

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    Area of Science:

    • Cell Biology
    • Biochemistry
    • Membrane Transport

    Background:

    • Lysosomes are key organelles involved in cellular degradation.
    • Understanding lysosomal membrane potential is critical for cellular processes.
    • The electrogenic nature of the lysosomal proton pump was previously unclear.

    Purpose of the Study:

    • To investigate the electrogenic properties of the lysosomal proton pump.
    • To quantify the lysosomal membrane potential (Δψ) using a fluorescent dye.
    • To elucidate the contribution of the proton gradient to Δψ.

    Main Methods:

    • Purification of rat kidney cortex lysosomes.
    • Utilized the potential-sensitive dye diS-C3-(5) to measure lysosomal membrane potential.
    • Employed ionophores (valinomycin, nigericin) and ion flux measurements (⁸⁶Rb⁺, ¹⁴C-SCN⁻) to validate findings.

    Main Results:

    • Lysosomes quenched diS-C3-(5) fluorescence, indicating dye accumulation and a measurable membrane potential.
    • Proton gradient collapse partially reversed fluorescence quenching, showing its contribution to Δψ.
    • Mg-ATP addition increased Δψ by 40-50 mV, demonstrating an electrogenic proton pump activity.

    Conclusions:

    • The lysosomal proton pump is electrogenic, actively contributing to the membrane potential.
    • ATP hydrolysis directly drives the generation of this electrical potential.
    • These findings provide crucial insights into lysosomal bioenergetics and function.

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