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Related Experiment Videos

Phosphoramidon-sensitive endothelin converting enzyme in rat lung.

M Takaoka1, K Shiragami, K Fujino

  • 1Department of Pharmacology, Osaka University of Pharmaceutical Sciences, Japan.

Biochemistry International
|November 1, 1991
PubMed
Summary

Researchers discovered a phosphoramidon-sensitive neutral proteinase in rat lungs that converts big endothelin-1 (ET-1) to ET-1. This enzyme may play a role in ET-1 production and regulation within the lung.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Physiology

Background:

  • Endothelin (ET)-1 is a potent vasoconstrictor peptide.
  • The conversion of big ET-1 to mature ET-1 is a critical regulatory step.
  • Understanding the enzymes involved in ET-1 processing is crucial for cardiovascular research.

Purpose of the Study:

  • To identify and characterize the proteinase responsible for big ET-1 conversion in rat lung.
  • To investigate the properties and potential role of this enzyme in ET-1 biosynthesis.

Main Methods:

  • Fractionation of rat lung cytosol and membranes.
  • Solubilization of membrane-bound proteinase using CHAPS.
  • Enzyme activity assays for big ET-1 conversion.
  • Inhibition studies with phosphoramidon.

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  • Molecular mass estimation by gel filtration.

    • Main Results:

    • A neutral proteinase with endothelin (ET)-1 converting activity was found in rat lung cytosol and membranes.
    • The membrane-bound enzyme was solubilized with CHAPS, showing increased specific activity.
    • The proteinase optimally converted big ET-1 to ET-1 at pH 6.5.
    • Phosphoramidon inhibited the conversion with an IC50 of 0.5 microM.
    • Gel filtration estimated the proteinase molecular mass at approximately 500 kDa.

    Conclusions:

    • Rat lungs possess a phosphoramidon-sensitive neutral proteinase that converts big ET-1 to ET-1.
    • This identified proteinase is likely involved in the biosynthetic pathway of ET-1 in the lung.
    • The enzyme may also contribute to the conversion of circulating big ET-1.