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Related Experiment Videos

ATP-dependent H+ pump in membrane vesicles from rat kidney cortex.

I Sabolić, W Haase, G Burckhardt

    The American Journal of Physiology
    |June 1, 1985
    PubMed
    Summary

    Researchers identified ATP-driven proton pumps in rat kidney endocytotic vesicles using acridine orange. These vesicles, distinct from other kidney cell membranes, play a role in proton transport within proximal tubule cells.

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    The Journal of chemical physics·2014

    Area of Science:

    • Nephrology
    • Cell Biology
    • Biochemistry

    Background:

    • The kidney cortex contains various membrane vesicles involved in transport processes.
    • Understanding the specific functions and origins of these vesicles is crucial for comprehending renal physiology.

    Purpose of the Study:

    • To identify and characterize ATP-driven proton (H+) pumps in rat kidney cortex homogenate.
    • To determine the cellular origin and specific characteristics of these H+ pump-containing vesicles.

    Main Methods:

    • Utilized the delta pH-sensitive dye acridine orange (AO) to detect H+ uptake.
    • Employed differential and Percoll density gradient centrifugation for vesicle purification.
    • Investigated vesicle identity using marker enzyme activities and in vivo labeling with horseradish peroxidase and FITC-dextran.

    Main Results:

    • Purified vesicles showed a 20-fold enrichment of ATP-driven H+ uptake compared to homogenate.
    • Vesicles were identified as endocytotic vesicles, distinct from brush border, basolateral membranes, lysosomes, and other organelles.
    • H+ pump activity in endocytotic vesicles was stimulated by chloride and inhibited by specific compounds like N-ethylmaleimide.

    Conclusions:

    • The study definitively identified endocytotic vesicles as containing an ATP-driven H+ pump in the rat kidney cortex.
    • These findings suggest that endocytotic vesicles contribute to proton transport mechanisms in proximal tubule cells.
    • The presence of similar H+ pumps in other membrane preparations may indicate contamination by endocytotic vesicles.

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