P2Y GPCR Assays
Background of P2Y Receptors
P2Y purinergic receptors are a family of extracellular G protein-coupled receptors for purine and pyrimidine nucleotides, such as ADP, ATP, and UTP, participating in purinergic signaling.
Fig.1 Structure of human P2Y1 receptor showing 7 transmembrane domains and intracellular domains for binding G proteins and protein kinases. (Zhang, 2015)
Distribution and Function of P2Y Receptors
P2Y receptors are mainly distributed in the placenta, prostate, brain, gastrointestinal tract, skeletal muscle, lung, heart, spleen, macrophages, lymphocytes, liver, and kidney. P2Y1 receptors participate in endothelial cell migration, mitogenic effects, smooth muscle relaxation, induction of platelet aggregation, regulation of EGF activity, and neuroprotection. P2Y2 receptors are involved in vasodilation, migration of vascular smooth muscle cells, orientation of neutrophil chemotaxis, and inhibition of apoptosis. P2Y4 receptors play a role in the activation of luminal K+ secretion, regulation of rod bipolar cell activity, adipogenesis and osteogenesis, etc. In addition, P2Y2 and P2Y4 receptors mediate the chloride epithelial transport in the jejunum. P2Y6 receptors are responsible for the contraction of smooth muscle, DNA synthesis, IL-8 release, Cl-secretion, tumor cell proliferation, regulation of insulin and glucagon secretion, etc. P2Y11 receptors play a role in the maturation and migration of DC cells, platelet release of nitric oxide, inhibition of neutrophil apoptosis, and chemotaxis of granulocytes. P2Y12 receptors are important in maintaining platelet aggregation and promoting thrombus growth and stabilization, as well as chemotaxis and response to arterial injury and thrombosis. P2Y13 receptors inhibit ATP release from erythrocytes, insulin secretion, and CFA-induced hyperalgesia. P2Y14 receptors are involved in chemotaxis, neuroimmune function, DC cell activation, etc.
Subtypes and Mechanisms of P2Y Receptors
The family of P2Y receptors in humans contains 8 subtypes of G protein-coupled receptors, including P2Y1, P2Y2, P2Y4, P2Y6, P2Y11, P2Y12, P2Y13, and P2Y14 receptors. They promote signal transduction primarily through heterotrimeric G proteins and ERK signaling pathways and regulation of ion channels, and kinases.
Receptor | Gene | Mechanism | Agonists | Antagonists |
P2Y1 receptor | P2RY1 |
|
|
|
P2Y2 receptor | P2RY2 |
|
|
|
P2Y4 receptor | P2RY4 |
|
|
|
P2Y6 receptor | P2RY6 |
|
|
|
P2Y11 receptor | P2RY11 |
|
|
|
P2Y12 receptor | P2RY12 |
|
|
|
P2Y13 receptor | P2RY13 |
|
|
|
P2Y14 receptor | P2RY14 |
|
|
|
Assay List of P2Y Receptors
Creative Biolabs can provide a range of assays of P2Y receptors. You can choose the assay in the list or contact us for more information:
Assay No. | Assay Name | Host Cell | Assay Type | Datasheet |
---|---|---|---|---|
IP1 Assay | ||||
S01YF-1122-KX845 | Magic™ Human P2RY1 In Vitro IP1 Assay | 1321N1 | IP1 Assay |
Assay No. | Assay Name | Host Cell | Assay Type | Datasheet |
---|---|---|---|---|
IP1 Assay | ||||
S01YF-1122-KX848 | Magic™ Human P2RY2 In Vitro IP1 Assay | 1321N1 | IP1 Assay |
Assay No. | Assay Name | Host Cell | Assay Type | Datasheet |
---|---|---|---|---|
IP1 Assay | ||||
S01YF-1122-KX849 | Magic™ Human P2RY4 In Vitro IP1 Assay | 1321N1 | IP1 Assay |
Published Data
Paper Title | Identification of P2Y receptors involved in oleamide-suppressing inflammatory responses in murine microglia and human dendritic cells |
Journal | Scientific reports |
Published | 2019 |
Abstract | The Previous study has found that oleamide is an endocannabinoid from fermented dairy products, which plays a role as a neuroprotective compound suppressing microglial inflammation. The suppressive effect of oleamide may be exerted via GPCR downstream signaling. Thus, the study aimed to explain the mechanism underlying the anti-inflammatory activity of the oleamide by screening the GPCRs using a transforming growth factor-α shedding assay system identified P2Y1, P2Y4, P2Y6, P2Y10, and P2Y11 as candidates for the oleamide target. They also observed a relationship between the P2Y1 agonistic activities and the suppressive activities of oleamide and its analogs. The study indicated that oleamide may exert as an antagonist for the P2Y1 receptor, which plays a role in the suppression of microglial inflammatory responses. |
Result |
The study has found that the suppressive effect of oleamide on microglial tumor necrosis factor-α (TNF-α) production was canceled by inhibitors of GPCR downstream signaling instead of a CB2 antagonist, which suggested that GPCRs are involved in the anti-inflammatory effects of oleamide. The result of the transforming growth factor-α shedding assay has demonstrated that the P2Y1 and P2Y10 agonists suppressed microglial TNF-α production, whereas a pan P2 receptor antagonist canceled the suppressive effect. The relationship between the P2Y1 agonistic activities and the suppressive activities of oleamide and its analogs, such as trans-OAD (tOAD), oleoylethylamide(OEtA), oleic acid, and palmitoylethanolamide (PEA). The analogs of oleamide displayed lower agonistic activity against P2Y1, P2Y4, and P2Y11 than oleamide, while oleoylethanolamide (OEA) had agonist activity. In summary, P2Y GPCRs are the potential targets of oleamide, and P2Y1 is important in the suppression of microglial inflammatory responses by oleamide.
Fig.2 Improvement in the TGF-α shedding response to OAD in HEK293 cells expressing P2Y1, P2Y4, P2Y6, P2Y10, P2Y11, CB1, and CB2. (Kita, 2019) |
References
- Zhang, D.; et al. Two disparate ligand-binding sites in the human P2Y1 receptor. Nature. 2015, 520(7547): 317-321.
- Kita, M.; et al. Identification of P2Y receptors involved in oleamide-suppressing inflammatory responses in murine microglia and human dendritic cells. Scientific reports. 2019, 9(1): 1-10.