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Related Experiment Videos

Coproporphyrinogen III oxidase assay

B Grandchamp, Y Nordmann

    Enzyme
    |January 1, 1982
    PubMed
    Summary

    A new assay measures coproporphyrinogen oxidase activity using a radiolabeled substrate. This sensitive and reproducible method quantifies enzyme activity in rat liver and human lymphocytes.

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    Area of Science:

    • Biochemistry
    • Enzymology
    • Molecular Biology

    Background:

    • Coproporphyrinogen oxidase (CPOX) is a key enzyme in heme biosynthesis.
    • Accurate measurement of CPOX activity is crucial for understanding related metabolic disorders.

    Purpose of the Study:

    • To develop a sensitive and reproducible assay for determining coproporphyrinogen oxidase activity.
    • To quantify CPOX activity in biological samples like rat liver and human lymphocytes.

    Main Methods:

    • Utilized a [14C]-coproporphyrinogen substrate for high specific activity.
    • Employed methylation, extraction, and thin-layer chromatography for product isolation.
    • Incorporated an internal standard to ensure assay reproducibility.

    Main Results:

    • The assay demonstrated high sensitivity and reproducibility.
    • Measured CPOX activity in rat liver as 140 nmol protoporphyrin/h/g.
    • Quantified CPOX activity in human lymphocytes as 483 pmol protoporphyrin/h/mg protein.

    Conclusions:

    • The developed assay is effective for measuring coproporphyrinogen oxidase activity.
    • Provides valuable quantitative data on CPOX levels in different tissues.
    • Facilitates further research into heme biosynthesis and associated pathologies.

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