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Related Experiment Videos

Phosphatidate phosphohydrolase and signal transduction

D N Brindley1, D W Waggoner

  • 1Signal Transduction Laboratory, Heritage Medical Research Centre, University of Alberta, Edmonton, Canada.

Chemistry and Physics of Lipids
|May 24, 1996
PubMed
Summary

A novel enzyme, phosphatidate phosphohydrolase-2 (PAP-2), regulates bioactive lipid signaling by dephosphorylating key substrates. PAP-2 influences lipid cross-talk, impacting cell activation pathways.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Lipid Metabolism

Background:

  • Bioactive lipids like phosphatidate, lysophosphatidate, sphingosine 1-phosphate, and ceramide 1-phosphate are critical signaling molecules.
  • These lipids play roles in various cellular processes, including cell activation and proliferation.
  • Dysregulation of these lipid signaling pathways is implicated in numerous diseases.

Purpose of the Study:

  • To identify and characterize a novel phosphatidate phosphohydrolase (PAP-2) involved in bioactive lipid metabolism.
  • To elucidate the enzymatic activity and substrate specificity of PAP-2.
  • To investigate the role of PAP-2 in the cross-talk between glycerolipid and sphingolipid signaling pathways.

Main Methods:

  • Purification of PAP-2 from cell plasma membranes.

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  • Enzymatic assays to determine substrate specificity and kinetics.
  • Inhibition studies using various lipid substrates and signaling molecules.
  • Analysis of lipid metabolite levels in response to enzyme activity and cellular stimulation.

    • Main Results:

    • A Mg(2+)-independent and N-ethylmaleimide-insensitive PAP-2 was purified.
    • PAP-2 dephosphorylates phosphatidate, lysophosphatidate, sphingosine 1-phosphate, and ceramide 1-phosphate.
    • Sphingolipids modulate PAP-2 activity and phosphatidate levels, indicating cross-talk between lipid pathways.
    • Ceramides inhibit phospholipase D and promote PAP-2 activity, further altering lipid balances.

    Conclusions:

    • PAP-2 is a key enzyme in terminating bioactive lipid signals and generating new signaling molecules.
    • PAP-2 plays a crucial role in integrating glycerolipid and sphingolipid signaling pathways.
    • Modulation of PAP-2 activity offers a potential target for regulating cellular responses to activation signals.