Competitive Antagonism of Recombinant P2X<sub>2/3</sub>Receptors by 2′,3′-<i>O</i>-(2,4,6-Trinitrophenyl) Adenosine 5′-Triphosphate (TNP-ATP)

Burgard, Edward C.; Niforatos, Wende; Biesen, Tim van; Lynch, Kevin J.; Kage, Karen; Touma, Edward; Kowaluk, Elizabeth A.; Jarvis, Michael F.

doi:10.1124/mol.58.6.1502

Cited by 66 publications

(69 citation statements)

References 32 publications

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“…5B). If TNP-ATP binds to the same site(s) as ATP, as is suggested for the homomeric P2X 3 receptor (Burgard et al, 2000) and for the homomeric P2X 2 receptor (Trujillo et al, 2006), then these results indicate that Lys 69 is involved in agonist binding.…”

Section: Partial Agonist Actions Of Suramin and Tnp-atp At P2x 2 [T339s]mentioning

confidence: 88%

Thr³³⁹-to-Serine Substitution in Rat P2X₂Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys³⁰⁸

Cao¹,

Young²,

Broomhead³

et al. 2007

J. Neurosci.

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Section: Partial Agonist Actions Of Suramin and Tnp-atp At P2x 2 [T339s]mentioning

confidence: 88%

Thr³³⁹-to-Serine Substitution in Rat P2X₂Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys³⁰⁸

Cao¹,

Young²,

Broomhead³

et al. 2007

J. Neurosci.

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“…The most potent competitive antagonist of P2X3 receptors is the 2′,3′-O-(2,4,6-trinitrophenyl)-ATP (TNP-ATP, Fig. 1) [47], with low nanomolar activity. In fact, this molecule showed to inhibit ATP or α,β-meATP evoked current at P2X3 receptors expressed by HEK293 cells with an IC 50 value of 2 nM [48].…”

Section: Introductionmentioning

confidence: 99%

2′,3′-O-Substituted ATP derivatives as potent antagonists of purinergic P2X3 receptors and potential analgesic agents

Ben

Marchenkova

Thomas

et al. 2016

Purinergic Signalling

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Blocking membrane currents evoked by the activation of purinergic P2X3 receptors localized on nociceptive neurons represents a promising strategy for the development of agents useful for the treatment of chronic pain conditions. Among compounds endowed with such antagonistic action, 2′,3′-O-(2,4,6-trinitrophenyl)-ATP (TNP-ATP) is an ATP analogue, whose inhibitory activity on P2X receptors has been previously reported. Based on the results of molecular modelling studies performed with homology models of the P2X3 receptor, novel adenosine nucleotide analogues bearing cycloalkyl or arylalkyl substituents replacing the trinitrophenyl moiety of TNP-ATP were designed and synthesized. These new compounds were functionally evaluated on native P2X3 receptors from mouse trigeminal ganglion (TG) sensory neurons using patch clamp recordings under voltage clamp configuration. Our data show that some of these molecules are potent (nanomolar range) and reversible inhibitors of P2X3 receptors, without any apparent effect on trigeminal GABA A and 5-HT 3 receptors, whose membrane currents were unaffected by the tested compounds.

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“…bound to trinitrophenyl ATP (TNP-ATP), which is composed of ATP and a trinitrophenyl group attached to the 2′ and 3′ hydroxyl of the ribose moiety (Fig. S6A) and is reportedly a representative non-subtype-selective antagonist of P2X receptors that competes with ATP (10,(39)(40)(41). We confirmed that the ATP-evoked currents of ΔzfP2X 4 -A′ were inhibited by TNP-ATP and that TNP-ATP did not evoke currents in oocytes expressing ΔzfP2X 4 -A′ (Fig.…”

Section: Nmr Resonances From Methionine Residues Introduced In Thementioning

confidence: 99%

Conductance of P2X ₄ purinergic receptor is determined by conformational equilibrium in the transmembrane region

Minato

Suzuki

Hara

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Ligand-gated ion channels are partially activated by their ligands, resulting in currents lower than the currents evoked by the physiological full agonists. In the case of P2X purinergic receptors, a cationselective pore in the transmembrane region expands upon ATP binding to the extracellular ATP-binding site, and the currents evoked by α,β-methylene ATP are lower than the currents evoked by ATP. However, the mechanism underlying the partial activation of the P2X receptors is unknown although the crystal structures of zebrafish P2X 4 receptor in the apo and ATP-bound states are available. Here, we observed the NMR signals from M339 and M351, which were introduced in the transmembrane region, and the endogenous alanine and methionine residues of the zebrafish P2X 4 purinergic receptor in the apo, ATP-bound, and α,β-methylene ATP-bound states. Our NMR analyses revealed that, in the α,β-methylene ATP-bound state, M339, M351, and the residues that connect the ATP-binding site and the transmembrane region, M325 and A330, exist in conformational equilibrium between closed and open conformations, with slower exchange rates than the chemical shift difference (<100 s −1 ), suggesting that the small population of the open conformation causes the partial activation in this state. Our NMR analyses also revealed that the transmembrane region adopts the open conformation in the state bound to the inhibitor trinitrophenyl-ATP, and thus the antagonism is due to the closure of ion pathways, except for the pore in the transmembrane region: i.e., the lateral cation access in the extracellular region.NMR | membrane proteins | ligand-gated ion channels | insect cell expression system | purinergic receptors I n chemical neurotransmission, various neurotransmitters bind to ligand-gated ion channels expressed in the plasma membrane of postsynaptic cells, such as the NMDA, AMPA, and P2X receptors, leading to changes in membrane potential and the concentration of intracellular ions. Each ligand for a ligand-gated ion channel has a distinct ability to evoke currents (1), and the ligands are classified according to the evoked current level: such as, full agonists, partial agonists, and antagonists. Partial agonists of ligand-gated ion channels reportedly offer clinical advantages over antagonists and full agonists in antidepressant and smoking-cessation treatment (2, 3).Two mechanisms have been proposed for the partial activation of the ligand-gated ion channels: the equilibrium between the open and closed conformations and the distinct conformation of the partial agonist-bound states from the closed and open conformations (4, 5). In the crystal structures of the extracellular region of the AMPA receptor, in which the distances between the two extracellular domains are changed upon agonist binding, the interdomain distances in the partial agonist-bound states correlated with the conductance level, suggesting that the AMPA receptor adopts specific intermediately permeable conformations (4, 6).The P2X receptors are a family of cation channe...

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Competitive Antagonism of Recombinant P2X_2/3Receptors by 2′,3′-O-(2,4,6-Trinitrophenyl) Adenosine 5′-Triphosphate (TNP-ATP)

Cited by 66 publications

References 32 publications

Thr³³⁹-to-Serine Substitution in Rat P2X₂Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys³⁰⁸

Thr³³⁹-to-Serine Substitution in Rat P2X₂Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys³⁰⁸

2′,3′-O-Substituted ATP derivatives as potent antagonists of purinergic P2X3 receptors and potential analgesic agents

Conductance of P2X ₄ purinergic receptor is determined by conformational equilibrium in the transmembrane region

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Competitive Antagonism of Recombinant P2X2/3Receptors by 2′,3′-O-(2,4,6-Trinitrophenyl) Adenosine 5′-Triphosphate (TNP-ATP)

Cited by 66 publications

References 32 publications

Thr339-to-Serine Substitution in Rat P2X2Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys308

Thr339-to-Serine Substitution in Rat P2X2Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys308

2′,3′-O-Substituted ATP derivatives as potent antagonists of purinergic P2X3 receptors and potential analgesic agents

Conductance of P2X 4 purinergic receptor is determined by conformational equilibrium in the transmembrane region

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Competitive Antagonism of Recombinant P2X_2/3Receptors by 2′,3′-O-(2,4,6-Trinitrophenyl) Adenosine 5′-Triphosphate (TNP-ATP)

Thr³³⁹-to-Serine Substitution in Rat P2X₂Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys³⁰⁸

Thr³³⁹-to-Serine Substitution in Rat P2X₂Receptor Second Transmembrane Domain Causes Constitutive Opening and Indicates a Gating Role for Lys³⁰⁸

Conductance of P2X ₄ purinergic receptor is determined by conformational equilibrium in the transmembrane region