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Creatine kinase activity in serum and uric acid solution

W A Warren, V Madelian

    Clinica Chimica Acta; International Journal of Clinical Chemistry
    |July 15, 1976
    PubMed
    Summary

    Skeletal muscle creatine kinases from humans and rabbits were inactivated by human serum and uric acid. Dithiothreitol fully reactivated enzymes from uric acid but only partially from serum.

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    Area of Science:

    • Biochemistry
    • Enzymology
    • Muscle Physiology

    Background:

    • Skeletal muscle creatine kinase (sCK) is crucial for cellular energy homeostasis.
    • Understanding sCK stability and inactivation mechanisms is vital for accurate diagnostic assays.
    • Serum components and metabolic byproducts can affect enzyme activity.

    Purpose of the Study:

    • To investigate the inactivation of human and rabbit skeletal muscle creatine kinases by human serum and uric acid.
    • To determine the efficacy of dithiothreitol (DTT) in reactivating inactivated sCK.
    • To assess the protective effect of DTT against sCK inactivation during storage.

    Main Methods:

    • Activation of human and rabbit sCK using thiol agents.
    • Incubation of activated sCK in human serum and uric acid solutions.
    • Measurement of enzyme activity before and after incubation.
    • Assessment of sCK reactivation using dithiothreitol.
    • Storage of serum samples with DTT at -17°C for 14 days.

    Main Results:

    • Both human and rabbit sCK were inactivated by human serum and uric acid.
    • sCK inactivated by uric acid was completely reactivated by DTT.
    • sCK inactivated by human serum showed only 60-70% reactivation by DTT.
    • No sCK inactivation was observed in serum samples containing 0.01 M DTT and stored at -17°C for 14 days.

    Conclusions:

    • Human serum contains factors that irreversibly inactivate skeletal muscle creatine kinase.
    • Dithiothreitol can fully reverse inactivation caused by uric acid but not by serum components.
    • Supplementation with DTT and cold storage effectively preserves sCK activity in serum samples.

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