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Procalcitonin is a glycoprotein

J W Jacobs, P K Lund, J T Potts

    The Journal of Biological Chemistry
    |March 25, 1981
    PubMed
    Summary
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    Messenger RNA from rat thyroid tumors directs the synthesis of a calcitonin precursor. This precursor undergoes glycosylation and cleavage within microsomal membranes, forming the mature, secreted hormone.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Endocrinology

    Background:

    • Calcitonin is a crucial hormone involved in calcium regulation.
    • Understanding the biosynthesis of calcitonin is essential for comprehending its physiological role and potential therapeutic applications.

    Purpose of the Study:

    • To investigate the co- and post-translational processing of the calcitonin precursor.
    • To elucidate the molecular mechanisms involved in the maturation and secretion of calcitonin.

    Main Methods:

    • Cell-free translation systems using mRNA from rat medullary carcinoma of the thyroid.
    • Incubation with canine pancreatic microsomal membranes.
    • Analysis of protein products using SDS-polyacrylamide gel electrophoresis, immunoprecipitation, concanavalin A-Sepharose binding, and enzymatic deglycosylation with endo-beta-N-acetylglucosaminidase H.

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  • Proteolytic digestion assays using trypsin and chymotrypsin.

    • Main Results:

    • A precursor of calcitonin (Mr = 15,000) was synthesized in vitro.
    • In the presence of microsomal membranes, a larger, membrane-processed precursor (apparent Mr = 17,000) was observed, which was immunoprecipitated by anti-calcitonin antiserum and bound to concanavalin A-Sepharose, indicating glycosylation.
    • Deglycosylation reduced the apparent molecular weight to 12,000.
    • The membrane-processed precursor was resistant to proteolytic digestion, unlike the initial precursor.

    Conclusions:

    • Calcitonin biosynthesis involves extensive co- and post-translational processing, including glycosylation and proteolytic cleavage of a precursor.
    • Microsomal membranes play a critical role in these processing events, including sequestration within vesicles.
    • The processed precursor is ultimately converted to the smaller, unglycosylated, secreted form of calcitonin.