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Related Experiment Videos

A radiochemical assay for heme synthase activity

J R Bloomer, K O Morton

    Enzyme
    |January 1, 1982
    PubMed
    Summary

    This study details a new radiochemical assay to measure heme synthase (ferrochelatase) activity. The method accurately quantifies low enzyme levels by tracking iron incorporation into heme.

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    Area of Science:

    • Biochemistry
    • Enzymology
    • Analytical Chemistry

    Background:

    • Heme synthesis is a critical metabolic pathway.
    • Accurate measurement of heme synthase (ferrochelatase) activity is essential for understanding heme biosynthesis disorders.
    • Existing methods may lack sensitivity for detecting low enzyme levels.

    Purpose of the Study:

    • To describe a novel radiochemical assay for quantifying heme synthase (ferrochelatase) activity.
    • To establish a sensitive method for detecting low levels of enzyme activity.

    Main Methods:

    • Utilizes the incorporation of radioisotope 59Fe into heme.
    • Maintains ferrous iron state and excludes oxygen during enzyme incubation.
    • Quantifies labeled heme product using gamma scintillation spectrometry after solvent extraction.

    Main Results:

    • The assay successfully measures the incorporation of 59Fe into heme.
    • The method is capable of detecting picomole amounts of heme product.
    • Demonstrates utility in situations with low heme synthase activity.

    Conclusions:

    • A sensitive and reliable radiochemical assay for heme synthase (ferrochelatase) activity has been developed.
    • This assay is particularly valuable for studying conditions with diminished enzyme function.
    • The method offers improved detection limits for biochemical analyses of heme metabolism.

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