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Cholecystokinin-converting enzymes in brain

A Malesci, E Straus, R S Yalow

    Proceedings of the National Academy of Sciences of the United States of America
    |January 1, 1980
    PubMed
    Summary

    Porcine brain extracts contain enzymes that process cholecystokinin (CCK) into smaller fragments like CCK-12 and CCK-8. Different enzymes, with distinct molecular sizes, cleave specific bonds within CCK.

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    Area of Science:

    • Biochemistry
    • Neuroscience
    • Enzymology

    Background:

    • Cholecystokinin (CCK) is a peptide hormone with diverse physiological roles.
    • The processing of CCK into biologically active fragments is crucial for its function.
    • Understanding the enzymes involved in CCK metabolism provides insights into neuroendocrine regulation.

    Purpose of the Study:

    • To investigate the enzymatic conversion of cholecystokinin (CCK) by porcine cerebral cortical extracts.
    • To identify and characterize the enzymes responsible for generating CCK fragments.
    • To compare the enzymatic activity with known proteases like trypsin.

    Main Methods:

    • Preparation of crude extracts from porcine cerebral cortical tissue.
    • Fractionation of extracts using Sephadex G-75 and G-50 gel filtration chromatography.
    • Analysis of cholecystokinin (CCK) cleavage products, specifically CCK-12 and CCK-8.
    • Determination of kinetic parameters (Km, Vmax) for enzymatic reactions.

    Main Results:

    • Crude porcine brain extracts convert CCK into dodecapeptide (CCK-12) and octapeptide (CCK-8) fragments.
    • A larger molecular radius enzyme (Sephadex G-75 eluate) cleaves the arginine-isoleucine bond, producing CCK-12.
    • A smaller molecular radius enzyme (Sephadex G-50 eluate) cleaves the arginine-aspartate bond, yielding both CCK-12 and CCK-8.
    • The brain enzyme exhibits a lower Km for CCK cleavage at the arginine-isoleucine bond compared to trypsin.

    Conclusions:

    • At least two distinct enzymes in porcine brain tissue are involved in CCK processing.
    • These enzymes differ in molecular size and cleavage specificity.
    • The characterized enzymes contribute to the generation of biologically relevant CCK fragments in the brain.

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