Caged agonist of P2Y1 and P2Y12 receptors for light-directed facilitation of platelet aggregation

Biochem Pharmacol. 2008 Mar 15;75(6):1341-7. doi: 10.1016/j.bcp.2007.10.037. Epub 2007 Dec 4.

Abstract

We have prepared a caged form (MRS2703) of a potent dual agonist of the P2Y(1) and P2Y(12) nucleotide receptors, 2-MeSADP, by blocking the beta-phosphate group with a 1-(3,4-dimethyloxyphenyl)eth-1-yl phosphoester. Although MRS2703 is itself inactive at human P2Y(1) and P2Y(12) receptors expressed heterologously in 1321N1 astrocytoma cells or in washed human platelets, this derivative readily regenerates the parent agonist upon mild irradiation with long-wave UV light (360 nm). The functional effect of the regenerated agonist was demonstrated by a rise in intracellular calcium mediated by either P2Y(1) or P2Y(12) receptors in transfected cells. Washed human platelets exposed to a solution of MRS2703 were induced to aggregate upon UV irradiation. At 1.0 microM MRS2703, full aggregation was achieved within 1 min of irradiation. Thus, this caged nucleotide promises to be a useful probe for potent P2Y receptor activation with light-directed spatial and temporal control.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Adenosine Diphosphate / analogs & derivatives*
  • Adenosine Diphosphate / chemical synthesis
  • Adenosine Diphosphate / pharmacology
  • Adenosine Diphosphate / radiation effects
  • Calcium / metabolism
  • Cell Line, Tumor
  • Cells, Cultured
  • Humans
  • Platelet Aggregation / drug effects*
  • Purinergic P2 Receptor Agonists*
  • Receptors, Purinergic P2 / genetics
  • Receptors, Purinergic P2Y1
  • Receptors, Purinergic P2Y12
  • Thionucleotides / pharmacology*
  • Thionucleotides / radiation effects
  • Transfection
  • Ultraviolet Rays

Substances

  • P1-2-(methylthio)adenosyl-P2-1-(4,5-dimethoxy-2-nitrophenyl)ethyl pyrophosphate
  • P2RY1 protein, human
  • P2RY12 protein, human
  • Purinergic P2 Receptor Agonists
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2Y1
  • Receptors, Purinergic P2Y12
  • Thionucleotides
  • methylthio-ADP
  • Adenosine Diphosphate
  • Calcium