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Caged cyclic ADP-ribose. Synthesis and use

R Aarhus1, K Gee, H C Lee

  • 1Department of Physiology, University of Minnesota, Minneapolis 55455.

The Journal of Biological Chemistry
|March 31, 1995
PubMed
Summary
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Researchers developed caged cyclic ADP-ribose (cADPR) for controlled calcium release. Photolysis with UV light effectively releases active cADPR, triggering calcium mobilization and egg activation.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Physiology

Background:

  • Cyclic ADP-ribose (cADPR) is a novel cyclic nucleotide.
  • cADPR plays a role in calcium (Ca2+) mobilization.

Purpose of the Study:

  • To synthesize and characterize a caged form of cADPR.
  • To investigate the controlled release and biological activity of cADPR.

Main Methods:

  • Synthesis of caged cADPR using 2-nitrophenethyldiazoethane.
  • Characterization by elemental analysis, 1H NMR, and HPLC.
  • Photolysis using UV light and spectrofluorimetry.
  • Microinjection into sea urchin eggs and observation of Ca2+ release and exocytosis.

Main Results:

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  • Caged cADPR was successfully synthesized and purified into two isomeric forms.
  • Both forms released active cADPR upon UV exposure (λ < 320 nm).
  • Photolyzed cADPR induced Ca2+ release in sea urchin egg homogenates, inhibited by 8-amino-cADPR.
  • Microinjected caged cADPR triggered Ca2+ mobilization and cortical exocytosis in live sea urchin eggs after photolysis.

    • Conclusions:

    • Caged cADPR provides a tool for light-induced, controlled release of active cADPR.
    • This method allows for precise temporal and spatial activation of Ca2+ signaling pathways.
    • Caged cADPR is effective in triggering biological responses, such as cortical exocytosis in sea urchin eggs.