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A specific 1,25-dihydroxyvitamin D3 binding macromolecule in chicken bone.

W S Mellon, H F DeLuca

    The Journal of Biological Chemistry
    |May 10, 1980
    PubMed
    Summary

    Researchers discovered a high-affinity macromolecule in chick bone cytosol that binds 1,25-dihydroxyvitamin D3. This molecule may act as a vitamin D receptor, crucial for bone physiology.

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    A new, highly sensitive assay for 1,25-dihydroxyvitamin D not requiring high-performance liquid chromatography: application of monoclonal antibody against vitamin D receptor to radioreceptor assay.

    Analytical biochemistry·1992

    Area of Science:

    • Endocrinology
    • Molecular Biology
    • Biochemistry

    Background:

    • Vitamin D is essential for calcium homeostasis and bone health.
    • The active form of vitamin D, 1,25-dihydroxyvitamin D3, exerts its effects by binding to specific receptors.
    • Understanding vitamin D metabolism and action in bone is critical for bone health.

    Purpose of the Study:

    • To identify and characterize macromolecules in chick bone cytosol that bind 1,25-dihydroxyvitamin D3.
    • To investigate the potential role of these binding molecules as receptors in bone.

    Main Methods:

    • Preparation of cytosol from vitamin D3-deficient chick bone homogenates.
    • Sucrose gradient centrifugation to analyze macromolecule sedimentation.
    • Radioligand binding assays using [26,27-3H]1,25-dihydroxyvitamin D3 to determine binding affinity and kinetics.
    • Competitive displacement assays with various vitamin D3 analogs.

    Main Results:

    • A 3.7 S macromolecule with high affinity (Kd = 7.6 x 10(-11) M) and low capacity for 1,25-dihydroxyvitamin D3 was identified.
    • This 3.7 S molecule is of intracellular origin and may aggregate under low salt conditions.
    • Several vitamin D3 metabolites, including 1 alpha-hydroxyvitamin D3 and 25-hydroxyvitamin D3, showed displacement activity, with 1,25-dihydroxyvitamin D3 being the most potent.
    • A separate 5.5 S molecule was observed, binding both 1,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3, with a preference for the latter.

    Conclusions:

    • Chick bone cytosol contains a specific, high-affinity binding macromolecule for 1,25-dihydroxyvitamin D3.
    • This macromolecule likely functions as a receptor, playing a role in the physiological actions of vitamin D in bone.

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