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Elastases from human and canine granulocytes, I. Some proteolytic and esterolytic properties.

K Ohlsson, I Olsson, M Delshammar

    Hoppe-Seyler'S Zeitschrift Fur Physiologische Chemie
    |September 1, 1976
    PubMed
    Summary
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    Human and canine granulocytes contain elastase enzymes. These enzymes exhibit similar substrate affinities to porcine pancreatic elastase, with Suc-Ala3-NHNp being the most convenient assay substrate.

    Area of Science:

    • Biochemistry
    • Enzymology
    • Cell Biology

    Background:

    • Human and canine granulocytes possess lysosome-like granules.
    • These granules contain enzymes exhibiting elastinolytic activity.

    Purpose of the Study:

    • To characterize the enzymatic behavior of granulocyte elastases.
    • To compare their substrate affinities with porcine pancreatic elastase.
    • To identify the optimal substrate for kinetic measurements and inhibition studies.

    Main Methods:

    • Enzymatic assays using protein substrates (elastin-orcein, azocasein) and synthetic substrates (Boc-Ala-ONp, Suc-Ala3-NHNp).
    • Photometric assays were employed for characterization.
    • Determination of dissociation constant (Ki) for human granulocyte elastase and human alpha1-antitrypsin complex.

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    Main Results:

    • Granulocyte elastases demonstrated significant elastinolytic activity.
    • Similar substrate affinities were observed between human/canine granulocyte elastases and porcine pancreatic elastase.
    • Suc-Ala3-NHNp was identified as the most suitable substrate for assaying human and dog granulocyte elastases.
    • The dissociation constant (Ki) for the human granulocyte elastase-alpha1-antitrypsin complex was determined as 3.5 x 10(-10)M.

    Conclusions:

    • Human and canine granulocyte elastases are characterized by their elastinolytic activity and substrate specificities.
    • Suc-Ala3-NHNp is a valuable tool for the kinetic analysis and inhibition studies of these enzymes.
    • The study provides crucial data on the interaction between human granulocyte elastase and its inhibitor, alpha1-antitrypsin.