Biochemical and Biophysical Research Communications
Identification of endogenous surrogate ligands for human P2Y12 receptors by in silico and in vitro methods☆
Section snippets
Methods
In silico screening. We used the AutoDock 3.0, a ligand flexible docking program [7], according to the manufacturer’s instructions with our in-house database. The number of grid points in the x-, y-, z-axis was 60 × 60 × 60 with grid points separated by 0.375 Å. The population size was set to 50. Each docking experiment consisted of a series of 200 simulations. The resulting initial set of 200 receptor–ligand configurations included crude configurations and required further refinement. To this end,
Results
The effects of the P2Y1 agonists and antagonists on the expressed receptor were measured using the cells stably expressing P2Y1 fusion receptors. First, the influences were investigated for 2-methylthio ADP (2MeSADP), which is a known P2Y1 agonist, on the [Ca2+]i of hP2Y1–hGqα and hP2Y1–hG16α cells (Fig. 1, upper). The results showed that 2MeSADP induced the [Ca2+]i increases in both of the cells with dose dependency. The EC50 values of 2MeSADP for the hP2Y1–Gqα and hP2Y1–hG16α receptors were
Discussion
GPCRs are integral membrane proteins and interact with endogenous ligands such as neurotransmitters and hormones. Takeda et al. [21] have analyzed the human genome sequences using SOSUI and have reported that about 950 kinds of GPCRs exist on the human genome. GPCRs are known to be the action sites of more than half of all contemporary medicines. However, since the membrane protein does not readily crystallize, very few of the tertiary structures of GPCRs have been clarified. On the other hand,
Acknowledgment
We thank Dr. E. Cooper in Ritsumeikan University for help in preparing the manuscript.
References (33)
- et al.
Identification of endogenous surrogate ligands for human P2Y receptors through an in silico search
J. Pharmacol. Sci.
(2004) - et al.
Characterization of the UDP-glucose receptor (re-named here the P2Y14 receptor) adds diversity to the P2Y receptor family
Trends Pharmacol. Sci.
(2003) - et al.
An expressed sequence tag (EST) data mining strategy succeeding in the discovery of new G-protein coupled receptors
J. Mol. Biol.
(2001) - et al.
Arachidonic acid metabolism in the human mast cell line HMC-1: 5-lipoxygenase gene expression and biosynthesis of thromboxane
Biochim. Biophys. Acta
(1995) - et al.
Leukotriene-receptor antagonist FPL-55712 and t-PA-induced thrombolysis in canine coronary thrombosis
Thromb. Res.
(1990) - et al.
Intravascular cysteinyl–leukotriene formation by clotting whole human blood. Evidence from clamped umbilical vein segments and thrombus specimens
Thromb. Res.
(1993) - et al.
Identification of G protein-coupled receptor genes from the human genome sequence
FEBS Lett.
(2002) - et al.
Receptor-Gα fusion proteins as a tool for ligand screening
Life Sci.
(2001) - et al.
G alpha 15 and G alpha 16 couple a wide variety of receptors to phospholipase C
J. Biol. Chem.
(1995) - et al.
Characterization of the human cysteinyl leukotriene 2 receptor
J. Biol. Chem.
(2000)
Characterization and cannel coupling of the P2Y12 nucleotide receptor of brain capillary endothelial cells
J. Biol. Chem.
A vision for the future of genomics research
Nature
Towards 3D structures of G protein-coupled receptors: multidisciplinary approach
Curr. Med. Chem.
ACoMFA analysis with conformational propensity: an attempt to analyze the SAR of a set of molecules with different conformational flexibility using a 3D-QSAR method
J. Comput. Aid. Mol. Des.
Construction of hypothetical three-dimensional structure of P2Y1 receptor based on Fourier transform analysis
J. Protein Chem.
Cited by (99)
The role of P2Y receptors in regulating immunity and metabolism
2021, Biochemical PharmacologyCitation Excerpt :Besides the naturally occurring nucleotide ligands of P2YRs, over the years several other agonists and antagonist, both natural and synthetic, have been identified (Table 1). In addition, non-nucleotide ligands have also been proposed, which include cysteinyl leukotriene E4 (P2Y12R), prostaglandin E2 glyceryl ester (P2Y6R) and 5-phosphoribosyl 1-pyrophosphate (P2Y1R and P2Y12R) [30,31]. This variety and flexibility in activation indicates the importance of P2YRs in signal transduction contributing to several biological pathways.
Pharmacological strategies for targeting platelet activation in asthma
2019, Current Opinion in PharmacologyThe leukotriene signaling pathway: a druggable target in Alzheimer's disease
2019, Drug Discovery TodayThe platelet P2 receptors
2019, PlateletsExpression and localization of GPR99 in human nasal mucosa
2017, Auris Nasus LarynxCitation Excerpt :This assumption is based on observations in CysLT1 and CysLT2 double-knockout mice where pro-inflammatory CysLT-mediated pathways persisted despite the loss of these receptors with ligand specificity for LTE4 [8]. An in silico study predicted that LTE4 might be a surrogate ligand for the purinergic P2Y12 receptor [9], and this receptor appears to be required for LTE4-mediated pulmonary inflammation [10]. Nevertheless, a recent report suggests that LTE4 does not bind directly to P2Y12 receptors, but acts as a co-receptor instead [11].
P2Y Receptors in Immune Response and Inflammation
2017, Advances in Immunology
- ☆
This study was partially supported by the Grant-in-Aid for scientific research from Japanese Ministry of Education, Culture, Sports, Science and Technology.
- 1
Present address: Department of Pharmacology, National Defense Medical College, Tokorozawa, Saitama 359-8513, Japan.