Identification of endogenous surrogate ligands for human P2Y12 receptors by in silico and in vitro methods

Biochem Biophys Res Commun. 2005 Nov 11;337(1):281-8. doi: 10.1016/j.bbrc.2005.09.052.


Endogenous ligands acting on a human P2Y12 receptor, one of the G-protein coupled receptors, were searched by in silico screening against our own database, which contains more than 500 animal metabolites. The in silico screening using the docking software AutoDock resulted in selection of cysteinylleukotrienes (CysLTs) and 5-phosphoribosyl 1-pyrophosphate (PRPP), with high free energy changes, in addition to the known P2Y12 ligands such as 2MeSADP and ADP. These candidates were subjected to an in vitro Ca2+ assay using the CHO cells stably expressing P2Y12-G16alpha fusion proteins. We found that CysLTE4 and PRPP acted on the P2Y12 receptor as agonists with the EC50 values of 1.3 and 7.8 nM, respectively. Furthermore, we analyzed the phylogenetic relationship of the P2Y, P2Y-like, and CysLT receptors based on sequence alignment followed by evolutionary analyses. The analyses showed that the P2Y12, P2Y13, P2Y14, GPR87, CysLT-1, and CysLT-2 receptors formed a P2Y-related receptor subfamily with common sequence motifs in the transmembrane regions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Animals
  • CHO Cells
  • Calcium / metabolism
  • Computational Biology
  • Cricetinae
  • Cricetulus
  • Humans
  • Leukotriene E4 / chemistry
  • Leukotriene E4 / pharmacology*
  • Leukotrienes / pharmacology
  • Ligands
  • Membrane Proteins / agonists*
  • Membrane Proteins / chemistry
  • Membrane Proteins / classification
  • Phosphoribosyl Pyrophosphate / chemistry
  • Phosphoribosyl Pyrophosphate / pharmacology*
  • Phylogeny
  • Purinergic P2 Receptor Agonists*
  • Receptors, Purinergic P2 / chemistry
  • Receptors, Purinergic P2 / classification
  • Receptors, Purinergic P2Y12


  • Leukotrienes
  • Ligands
  • Membrane Proteins
  • P2RY12 protein, human
  • Purinergic P2 Receptor Agonists
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2Y12
  • Phosphoribosyl Pyrophosphate
  • Leukotriene E4
  • Calcium