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Related Experiment Videos

Cell-specific differences in membrane beta-glucosidase from normal and Gaucher cells.

B M Turner, N G Beratis, K Hirschhorn

    Biochimica Et Biophysica Acta
    |February 9, 1977
    PubMed
    Summary

    Researchers identified two human cell beta-glucosidase isozymes: acid-labile and acid-stable. Gaucher

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    Area of Science:

    • Biochemistry
    • Enzymology
    • Cell Biology

    Background:

    • Human cells possess membrane-bound beta-glucosidase (beta-D-glucoside glucohydrolase, EC 3.2.1.21) involved in cellular processes.
    • Two distinct isozymes of this enzyme, differing in pH stability and kinetic properties, have been previously suggested.

    Purpose of the Study:

    • To characterize the biochemical properties of two human beta-glucosidase isozymes.
    • To investigate the distribution of these isozymes across different human cell types.
    • To examine the impact of Gaucher's disease on the activity of these beta-glucosidase isozymes.

    Main Methods:

    • Enzyme activity assays using 4-methylumbelliferyl-beta-D-glucopyranoside as a substrate.
    • Determination of pH optima, Michaelis constants (Km), and stability profiles.
    • Analysis of enzyme activity in cultured lymphoid lines, peripheral blood leukocytes, and skin fibroblasts.
    • Enzyme solubilization studies using Triton X-100 and characterization of solubilized enzyme properties.

    Main Results:

    • Two isozymes were identified: an 'acid-labile' form (pH optimum 5.0, Km 0.4 mM) and an 'acid-stable' form (pH optimum 4.5, Km 0.8 mM).
    • Lymphoid lines and leukocytes contained both isozymes, while fibroblasts predominantly showed the 'acid-stable' form.
    • Gaucher's disease significantly reduced 'acid-stable' isozyme activity in all cell types, while 'acid-labile' activity remained unaffected in lymphoid cells and leukocytes.
    • Solubilized beta-glucosidase, requiring sodium taurocholate, lost cell-specific differences in pH optimum and Km.

    Conclusions:

    • Distinct biochemical properties and cellular distributions characterize the two beta-glucosidase isozymes.
    • The 'acid-stable' isozyme is implicated in Gaucher's disease pathology, showing reduced activity in affected cells.
    • Enzyme solubilization alters kinetic properties, suggesting membrane association influences enzyme behavior.

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