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Related Experiment Videos

Aldosterone-induced proteins: characterization using lectin-affinity chromatography.

B Blazer-Yost, M Cox

    The American Journal of Physiology
    |September 1, 1985
    PubMed
    Summary
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    Aldosterone-induced proteins (AIPs) are crucial for sodium transport in toad bladders. These proteins are N-linked glycoproteins, and differences in their glycosylation explain their varied molecular weights and aid in purification.

    Area of Science:

    • Molecular Endocrinology
    • Cellular Physiology
    • Glycobiology

    Background:

    • Aldosterone stimulates Na+ transport in toad urinary bladders via aldosterone-induced proteins (AIPs).
    • AIPs exhibit heterogeneity in molecular weight and isoelectric points, suggesting post-translational modifications like glycosylation.
    • Understanding the role of glycosylation in AIP function is key to elucidating aldosterone's mechanism of action.

    Purpose of the Study:

    • To investigate whether aldosterone-induced proteins (AIPs) are glycoproteins.
    • To determine the role of glycosylation in aldosterone-stimulated Na+ transport and AIP synthesis.
    • To characterize the N-linked glycosylation of AIPs and explore their purification using lectin affinity chromatography.

    Main Methods:

    • Inhibition of N-linked glycosylation using tunicamycin.

    Related Experiment Videos

  • Analysis of aldosterone-stimulated Na+ transport and protein synthesis.
  • Concanavalin A-sepharose affinity chromatography to assess lectin binding and elution profiles of AIPs.

    • Main Results:

    • Tunicamycin treatment inhibited both aldosterone-stimulated Na+ transport and AIP synthesis.
    • Aldosterone-induced proteins (AIPs) bound to concanavalin A-sepharose, indicating mannose-containing N-linked glycosylation.
    • Differential elution of AIPs from concanavalin A-sepharose suggested variations in glycosylation patterns between molecular weight classes.

    Conclusions:

    • Glycosylation is essential for the natriferic response to aldosterone.
    • Aldosterone-induced proteins (AIPs) are N-linked mannose-containing glycoproteins.
    • Differences in glycosylation contribute to the electrophoretic polymorphism of AIPs, and concanavalin A chromatography aids in their purification.