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Related Experiment Videos

Cytosolic and membrane-bound nitric oxide synthase.

K Hiki1, Y Yui, R Hattori

  • 1Department of Internal Medicine, Faculty of Medicine, Kyoto University, Japan.

Japanese Journal of Pharmacology
|June 1, 1991
PubMed
Summary

This study reveals two forms of nitric oxide (NO) synthase in macrophages: a cytosolic form and a membrane-bound enzyme. These enzymes exhibit distinct calcium dependencies, influencing NO production.

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Area of Science:

  • Biochemistry
  • Immunology
  • Cell Biology

Background:

  • Macrophages play a crucial role in immune responses.
  • Nitric oxide (NO) is a key signaling molecule produced by macrophages.
  • The regulation and localization of NO synthase (NOS) enzymes are critical for controlling NO production.

Purpose of the Study:

  • To investigate the different forms and localization of NO synthase (NOS) in cytotoxic activated macrophages.
  • To determine the calcium dependency of NOS activity in different cellular fractions.

Main Methods:

  • Macrophages were sonicated and centrifuged to separate cellular components.
  • Supernatants and pellets were analyzed for NO synthase activity.
  • Differential extraction using Triton X-100 was employed to solubilize membrane-bound proteins.

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  • Calcium dependency of NO synthase activity was assessed.

    • Main Results:

    • NO synthase activity was initially found in the supernatant, independent of calcium.
    • After Triton X-100 treatment, a calcium-dependent NO synthase activity was recovered.
    • The pellet fraction, after detergent treatment, revealed a distinct calcium-dependent NOS activity.

    Conclusions:

    • These findings suggest the presence of at least two distinct forms of NO synthase in macrophages.
    • A cytosolic, calcium-independent NOS and a membrane-bound, calcium-dependent NOS are proposed.
    • Understanding these distinct NOS forms is crucial for regulating macrophage-mediated immune responses and NO signaling.