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Platelet and leucocyte calmodulins: isolation and characterisation.

N Young, P Gergely, N Crawford

    European Journal of Biochemistry
    |November 1, 1981
    PubMed
    Summary

    Researchers isolated and purified calmodulins from human blood platelets and guinea pig polymorphonuclear leucocytes. These proteins demonstrated significant biological activity, stimulating key enzymes involved in cellular regulation.

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    Area of Science:

    • Biochemistry
    • Cell Biology
    • Protein Chemistry

    Background:

    • Calmodulins are crucial calcium-dependent regulatory proteins involved in various cellular processes.
    • Previous research has established the presence and function of calmodulins in diverse mammalian tissues.

    Purpose of the Study:

    • To isolate and characterize calmodulins from human blood platelets and guinea pig polymorphonuclear leucocytes.
    • To compare the yield and biological activities of these newly isolated calmodulins with known calmodulin preparations.

    Main Methods:

    • Proteins were isolated using the urea methanol procedure.
    • Purification to homogeneity was confirmed by polyacrylamide gel electrophoresis.
    • Molecular weights were determined by gel electrophoresis, and biological activity was assessed by enzyme stimulation assays.

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    Main Results:

    • Calmodulins were successfully isolated and purified from both cell types, exhibiting molecular weights of 16,000-17,000.
    • Platelet calmodulin yielded higher amounts compared to leucocyte calmodulin, comparable to brain tissue.
    • Both preparations significantly stimulated cyclic nucleotide phosphodiesterase, Ca2+ ATPase, and phosphorylase kinase activities.

    Conclusions:

    • Human blood platelets and guinea pig polymorphonuclear leucocytes are viable sources for calmodulin isolation.
    • The isolated calmodulins possess significant biological activities, confirming their regulatory roles.
    • While stable during storage, aggregate formation was observed, suggesting potential long-term stability issues.