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Related Experiment Videos

Rabbit heart mitochondrial hexokinase: solubilization and general properties.

E Aubert-Foucher, B Font, D C Gautheron

    Archives of Biochemistry and Biophysics
    |July 1, 1984
    PubMed
    Summary

    Rabbit heart mitochondria contain hexokinase type I (HK I). Specific interactions with the outer mitochondrial membrane are mediated by glucose 6-phosphate and cation-dependent binding, influencing enzyme activity.

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    Area of Science:

    • Biochemistry
    • Cell Biology
    • Enzymology

    Background:

    • Hexokinase (HK) is a key enzyme in glucose metabolism, primarily found in the mitochondria.
    • Rabbit heart tissue exhibits two HK isoenzymes, with mitochondrial activity attributed to HK type I (HK I).

    Purpose of the Study:

    • To investigate the interactions between rabbit heart hexokinase and the mitochondrial membrane.
    • To characterize the binding and solubilization properties of mitochondrial-bound HK I.

    Main Methods:

    • Enzyme kinetics (apparent Km determination for glucose and ATPMg).
    • Electrophoresis to identify isoenzymes.
    • Differential solubilization using neutral salts, digitonin, glucose 6-phosphate (G6P), and ATP.
    • Inhibition studies with Pi and Mg2+.

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    Main Results:

    • Mitochondrial HK is exclusively HK I, with specific kinetic properties (Km for glucose = 0.065 mM).
    • Bound HK I shows altered ATPMg kinetics compared to solubilized enzyme.
    • Specific solubilization observed with low concentrations of G6P or ATP4-.
    • Solubilization is modulated by Pi, Mg2+, and the integrity of mitochondrial membranes.

    Conclusions:

    • Rabbit heart HK I interacts with the outer mitochondrial membrane via specific and cation-mediated interactions.
    • These interactions are analogous to models proposed for brain hexokinase.
    • G6P and ATP4- act as specific solubilizing agents, suggesting a regulatory role in enzyme localization and activity.