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Sulfite oxidase from Merluccius productus.

Y Onoue

    Biochimica Et Biophysica Acta
    |September 9, 1980
    PubMed
    Summary

    Pacific hake sulfite oxidase was purified and characterized. This enzyme, crucial for sulfite metabolism, was found to be a dimeric protein containing molybdenum and heme, primarily located in liver mitochondria.

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    Area of Science:

    • Biochemistry
    • Enzymology
    • Marine Biology

    Background:

    • Sulfite oxidase (EC 1.8.3.1) is a key enzyme in sulfur metabolism.
    • Understanding its properties in marine species provides comparative biochemical insights.

    Purpose of the Study:

    • To purify and characterize sulfite oxidase from the Pacific hake (Merluccius productus).
    • To determine the enzyme's molecular properties, kinetic parameters, and cofactor content.
    • To investigate the subcellular localization and tissue distribution of sulfite oxidase activity.

    Main Methods:

    • Enzyme purification using chromatography (Sephadex G-100).
    • Molecular weight determination via gel exclusion chromatography and SDS-PAGE.
    • Enzyme kinetics measured using sulfite and cytochrome c as substrates.
    • Spectroscopic analysis (EPR) and colorimetric assays for cofactor identification.
    • Differential centrifugation for subcellular localization studies.

    Main Results:

    • Sulfite oxidase was purified 482-fold from Pacific hake liver.
    • The enzyme has a molecular weight of 120,000 Da, composed of two 60,000 Da subunits.
    • Optimal activity at pH 8.7, with Ks values for sulfite and cytochrome c determined.
    • EPR and colorimetric analyses confirmed the presence of pentavalent molybdenum and heme.
    • Mitochondria contained the majority of sulfite oxidase activity.
    • Highest activity observed in liver and kidney tissues.

    Conclusions:

    • The purified sulfite oxidase from Pacific hake exhibits characteristics consistent with mammalian sulfite oxidases.
    • The enzyme is a dimeric metalloprotein containing molybdenum and heme, localized primarily in mitochondria.
    • Tissue distribution suggests a significant role in sulfite detoxification in the liver and kidney of this marine species.

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