Abstract
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 Ca 2+ assay using the CHO cells stably expressing P2Y 12-G16α 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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 281-288 |
| Number of pages | 8 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 337 |
| Issue number | 1 |
| DOIs | |
| State | Published - Nov 11 2005 |
| Externally published | Yes |
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Keywords
- 3D model of GPCR
- 5-phosphoribosyl 3-pyrophosphate
- Cysteinylleukotriene E4
- Gα-mediated Ca response
- In silico screening
- P2Y receptor
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Identification of endogenous surrogate ligands for human P2Y12 receptors by in silico and in vitro methods. / Nonaka, Yosuke; Hiramoto, Takeshi; Fujita, Norihisa.
In: Biochemical and Biophysical Research Communications, Vol. 337, No. 1, 11.11.2005, p. 281-288.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of endogenous surrogate ligands for human P2Y12 receptors by in silico and in vitro methods
AU - Nonaka, Yosuke
AU - Hiramoto, Takeshi
AU - Fujita, Norihisa
PY - 2005/11/11
Y1 - 2005/11/11
N2 - 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 Ca 2+ assay using the CHO cells stably expressing P2Y 12-G16α 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.
AB - 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 Ca 2+ assay using the CHO cells stably expressing P2Y 12-G16α 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.
KW - 3D model of GPCR
KW - 5-phosphoribosyl 3-pyrophosphate
KW - Cysteinylleukotriene E4
KW - Gα-mediated Ca response
KW - In silico screening
KW - P2Y receptor
UR - http://www.scopus.com/inward/record.url?scp=25844519676&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=25844519676&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2005.09.052
DO - 10.1016/j.bbrc.2005.09.052
M3 - Article
VL - 337
SP - 281
EP - 288
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -