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Cutaneous alkaline phosphatase: a biochemical study

P D Mier, H van Rennes

    Archives of Dermatological Research
    |January 1, 1982
    PubMed
    Summary
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    A new fluorometric method offers highly sensitive alkaline phosphatase detection in skin. This assay, requiring fluoride, reveals the enzyme is primarily in the dermis, not epidermis.

    Area of Science:

    • Biochemistry
    • Dermatology
    • Enzymology

    Background:

    • Alkaline phosphatase (ALP) is crucial in various physiological processes.
    • Existing colorimetric assays for ALP lack sensitivity.
    • Skin ALP activity is poorly characterized, with potential epidermal interference.

    Purpose of the Study:

    • To develop and validate a highly sensitive fluorometric microassay for alkaline phosphatase.
    • To investigate the kinetic properties of alkaline phosphatase in human skin.
    • To determine the localization and forms of alkaline phosphatase within skin tissues.

    Main Methods:

    • Fluorometric microassay using 4-methylumbelliferyl phosphate as substrate.
    • Application of the assay to human skin samples.

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  • Inclusion of fluoride to mitigate acid phosphatase interference.
  • Kinetic analysis of enzyme activity.

    • Main Results:

    • The fluorometric assay is approximately 1000 times more sensitive than conventional colorimetric methods.
    • Fluoride addition is essential for accurate measurements in skin due to high epidermal acid phosphatase.
    • Cutaneous alkaline phosphatase exhibits kinetics characteristic of the 'bone-liver-kidney' isoenzyme.
    • The enzyme exists in both soluble and membrane-bound forms within the skin.
    • Alkaline phosphatase is predominantly located in the dermis, with less than 1% in the epidermis.

    Conclusions:

    • A sensitive fluorometric assay for skin alkaline phosphatase has been established.
    • Human skin contains a single type of alkaline phosphatase, primarily localized in the dermis.
    • Understanding skin ALP distribution is vital for dermatological research and diagnostics.