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Title: Identification of endogenous surrogate ligands for human P2Y{sub 12} receptors by in silico and in vitro methods

Abstract

Endogenous ligands acting on a human P2Y{sub 12} 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 P2Y{sub 12} ligands such as 2MeSADP and ADP. These candidates were subjected to an in vitro Ca{sup 2+} assay using the CHO cells stably expressing P2Y{sub 12}-G{sub 16}{alpha} fusion proteins. We found that CysLTE4 and PRPP acted on the P2Y{sub 12} receptor as agonists with the EC{sub 50} 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 P2Y{sub 12}, P2Y{sub 13}, P2Y{sub 14}, GPR87, CysLT-1, and CysLT-2 receptors formed a P2Y-related receptor subfamily with common sequence motifs in the transmembrane regions.

Authors:
 [1];  [1];  [2]
  1. Laboratory of Pharmcoinformatics, Department of Bioinformatics, Ritsumeikan University, Kusatsu, Shiga 525-8577 (Japan)
  2. Laboratory of Pharmcoinformatics, Department of Bioinformatics, Ritsumeikan University, Kusatsu, Shiga 525-8577 (Japan). E-mail: nori@is.ritsumei.ac.jp
Publication Date:
OSTI Identifier:
20713436
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 337; Journal Issue: 1; Other Information: DOI: 10.1016/j.bbrc.2005.09.052; PII: S0006-291X(05)02061-9; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ADP; ALIGNMENT; CALCIUM IONS; CHO CELLS; COMPUTER CODES; FREE ENERGY; GTP-ASES; IN VITRO; LIGANDS; RECEPTORS

Citation Formats

Nonaka, Yosuke, Hiramoto, Takeshi, and Fujita, Norihisa. Identification of endogenous surrogate ligands for human P2Y{sub 12} receptors by in silico and in vitro methods. United States: N. p., 2005. Web. doi:10.1016/j.bbrc.2005.09.052.
Nonaka, Yosuke, Hiramoto, Takeshi, & Fujita, Norihisa. Identification of endogenous surrogate ligands for human P2Y{sub 12} receptors by in silico and in vitro methods. United States. doi:10.1016/j.bbrc.2005.09.052.
Nonaka, Yosuke, Hiramoto, Takeshi, and Fujita, Norihisa. Fri . "Identification of endogenous surrogate ligands for human P2Y{sub 12} receptors by in silico and in vitro methods". United States. doi:10.1016/j.bbrc.2005.09.052.
@article{osti_20713436,
title = {Identification of endogenous surrogate ligands for human P2Y{sub 12} receptors by in silico and in vitro methods},
author = {Nonaka, Yosuke and Hiramoto, Takeshi and Fujita, Norihisa},
abstractNote = {Endogenous ligands acting on a human P2Y{sub 12} 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 P2Y{sub 12} ligands such as 2MeSADP and ADP. These candidates were subjected to an in vitro Ca{sup 2+} assay using the CHO cells stably expressing P2Y{sub 12}-G{sub 16}{alpha} fusion proteins. We found that CysLTE4 and PRPP acted on the P2Y{sub 12} receptor as agonists with the EC{sub 50} 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 P2Y{sub 12}, P2Y{sub 13}, P2Y{sub 14}, GPR87, CysLT-1, and CysLT-2 receptors formed a P2Y-related receptor subfamily with common sequence motifs in the transmembrane regions.},
doi = {10.1016/j.bbrc.2005.09.052},
journal = {Biochemical and Biophysical Research Communications},
number = 1,
volume = 337,
place = {United States},
year = {Fri Nov 11 00:00:00 EST 2005},
month = {Fri Nov 11 00:00:00 EST 2005}
}