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The contribution of the intracellular domains to P2X1 receptor regulation

By Hairuo Wen


P2X1 receptors are expressed throughout the body and contribute to a range of physiological process, e.g. thrombosis and smooth muscle tone. The intracellular terminals of the P2X1 receptor have been shown to be involved in channel regulation.\ud The aim of this thesis was to explore the contribution of the intracellular amino and carboxy termini to channel regulation, using concatenation, site-directed mutagenesis and biochemical methods. Trimeric concatenated P2X1 receptors were non-functional.The interactions between the intracellular termini were revealed to be essential for subunit aggregation and channel expression. Subsequently, over 30 cysteine point mutations at both the amino and carboxy domains of the P2X1 receptors were generated and examined by electrophysiological studies with methanethiosulfonate modifications. The conserved TXTXK/R at the amino terminal and the residues around the YXXXK motif at the carboxy terminal were highlighted to be important for channel gating and protein trafficking. Residues close to the P2X1 channel pore were shown to be essential in ion conduction and implied interactions between the conserved regulatory motifs and the channel pore of adjacent residues.\ud Roles of the intracellular domains of P2X1 receptor in regulation by protein kinase were revealed by co-expression of minigenes encoding the sequences of amino/carboxy terminus with WT receptors. The results indicated both amino and carboxy termini contribute to the phorbol ester PMA and GPCR mediated response regulations, and demonstrated the important roles of the conserved TXTXK/R motif and the residues around the YXXXK motif. This research sheds light on the possible interactions between the amino and carboxy termini to P2X1 receptor function. The results will be useful for understanding the intracellular topology and signaling modulation mechanism of P2X1 receptor

Publisher: University of Leicester
Year: 2009
OAI identifier: oai:lra.le.ac.uk:2381/9054

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  1. (1958). [Noradrenalin and ATP content of sympathetic nerves.].
  2. (1978). A basis for distinguishing two types of purinergic receptors,
  3. (2006). A C-terminal lysine that controls human P2X4 receptor desensitization.
  4. (1998). A domain contributing to the ion channel of ATP-gated P2X2 receptors identified by the substituted cysteine accessibility method.
  5. (2000). A G protein-coupled receptor for UDP-glucose.
  6. (1963). A Molecular Theory of Muscle Contraction: Calcium-Dependent Contractions with Hydrogen Bond Formation Plus Atp-Dependent Extensions of Part of the Myosin-Actin Cross-Bridges.
  7. (1997). A mutational analysis of residues essential for ligand recognition at the human P2Y1 receptor.
  8. (1994). A new class of ligand-gated ion channel defined by P2x receptor for extracellular ATP.
  9. (1987). A novel receptor-operated Ca2+-permeable channel activated by ATP in smooth muscle.
  10. (1995). A P2X purinoceptor cDNA conferring a novel pharmacological profile.
  11. (1995). A P2X purinoceptor expressed by a subset of sensory neurons.
  12. (2000). A protein kinase C site highly conserved in P2X subunits controls the desensitization kinetics of P2X(2) ATP-gated channels.
  13. (1998). A pyridoxine cyclic phosphate and its 6-azoaryl derivative selectively potentiate and antagonize activation of P2X1 receptors.
  14. (1997). A role for calcineurin in the desensitization of the P2X3 receptor.
  15. (2003). A role of the fast ATP-gated P2X1 cation channel in thrombosis of small arteries in vivo.
  16. (2002). A study of P2X1 receptor function in murine megakaryocytes and human platelets reveals synergy with P2Y receptors.
  17. (2006). A truncated P2X7 receptor variant (P2X7-j) endogenously expressed in cervical cancer cells antagonizes the full-length P2X7 receptor through hetero-oligomerization.
  18. (2002). A-317491, a novel potent and selective non-nucleotide antagonist of P2X3 and P2X2/3 receptors, reduces chronic inflammatory and neuropathic pain in the rat.
  19. (2002). Absence of the P2X7 receptor alters leukocyte function and attenuates an inflammatory response.
  20. (1990). Acetylcholine and ATP are coreleased from the electromotor nerve terminals of Narcine brasiliensis by an exocytotic mechanism.
  21. (1992). Acetylcholine receptor channel structure probed in cysteine-substitution mutants.
  22. (2003). Activation and desensitization of the recombinant P2X1 receptor at nanomolar ATP concentrations.
  23. (2001). Altered cytokine production in mice lacking P2X(7) receptors.
  24. (2001). Amino acid residues involved in gating identified in the first membrane-spanning domain of the rat P2X(2) receptor.
  25. (1993). Amino acids lining the channel of the gamma-aminobutyric acid type A receptor identified by cysteine substitution.
  26. (2003). An Ile-568 to Asn polymorphism prevents normal trafficking and function of the human P2X7 receptor.
  27. (2005). An intersubunit zinc binding site in rat P2X2 receptors.
  28. (2007). An intracellular motif of P2X(3) receptors is required for functional cross-talk with GABA(A) receptors in nociceptive DRG neurons.
  29. (2007). An intracellular P2X receptor required for osmoregulation in Dictyostelium discoideum.
  30. (2006). An uncharged region within the N terminus of the P2X6 receptor inhibits its assembly and exit from the endoplasmic reticulum.
  31. (1999). Antagonistic properties of the suramin analogue NF023 at heterologously expressed P2X receptors.
  32. (2000). Asparagine-mediated self-association of a model transmembrane helix.
  33. (2002). Assembly and cell surface expression of homomeric and heteromeric 5-HT3 receptors: the role of oligomerization and chaperone proteins.
  34. (2005). Atomic force microscopy imaging demonstrates that P2X2 receptors are trimers but that P2X6 receptor subunits do not oligomerize.
  35. (2004). ATP binding at human P2X1 receptors. Contribution of aromatic and basic amino acids revealed using mutagenesis and partial agonists.
  36. (2002). ATP binding to the motor domain from an ABC transporter drives formation of a nucleotide sandwich dimer.
  37. (1972). Atropine resistant excitation of the urinary bladder: the possibility of transmission via nerves releasing a purine nucleotide.
  38. (1997). Baculovirus expression provides direct evidence for heteromeric assembly of P2X2 and P2X3 receptors.
  39. (2005). Biochemical and functional evidence for heteromeric assembly of P2X1 and P2X4 subunits.
  40. (2005). Biochemistry and molecular biology, Italy,
  41. (1993). Calcineurin inhibits desensitization of cloned rat 5-HT1C receptors.
  42. (1998). Calcium permeability and block at homomeric and heteromeric P2X2 and P2X3 receptors, and P2X receptors in rat nodose neurones.
  43. (1998). Central P2X4 and P2X6 channel subunits coassemble into a novel heteromeric ATP receptor.
  44. (1994). Characterization of P2-purinoceptors in the smooth muscle of the rat tail artery: a comparison between contractile and electrophysiological responses.
  45. (2003). Characterization of the UDP-glucose receptor (re-named here the P2Y14 receptor) adds diversity to the P2Y receptor family.
  46. (1997). Cloned ligand-gated channels activated by extracellular ATP (P2X receptors).
  47. (1993). Cloning and functional expression of a brain G-protein-coupled ATP receptor.
  48. (1996). Cloning OF P2X5 and P2X6 receptors and the distribution and properties of an extended family of ATP-gated ion channels.
  49. (2005). Clopidogrel and ticlopidine: P2Y12 adenosine diphosphate-receptor antagonists for the prevention of atherothrombosis.
  50. (1998). Co-expression of P2X1 and P2X5 receptor subunits reveals a novel ATP-gated ion channel.
  51. (1995). Coexpression of P2X2 and P2X3 receptor subunits can account for ATP-gated currents in sensory neurons.
  52. (2002). Conserved cysteine residues in the extracellular loop of the human P2X(1) receptor form disulfide bonds and are involved in receptor trafficking to the cell surface.
  53. (2001). Conserved negatively charged residues are not required for ATP action at P2X(1) receptors.
  54. (2004). Contribution of calcium ions to P2X channel responses.
  55. (2005). Contribution of conserved glycine residues to ATP action at human P2X1 receptors: mutagenesis indicates that the glycine at position 250 is important for channel function.
  56. (2006). Contribution of conserved polar glutamine, asparagine and threonine residues and glycosylation to agonist action at human P2X1 receptors for ATP.
  57. (1999). Contribution of individual subunits to the multimeric P2X(2) receptor: estimates based on methanethiosulfonate block at T336C.
  58. (2006). Contribution of P2X1 receptor intracellular basic residues to channel properties.
  59. (2002). Control of P2X(2) channel permeability by the cytosolic domain.
  60. (2004). Cross-talk and co-trafficking between rho1/GABA receptors and ATP-gated channels.
  61. (2008). Cysteine substitution mutagenesis and the effects of methanethiosulfonate reagents at P2X2 and P2X4 receptors support a core common mode of ATP action at P2X receptors.
  62. (2007). Cysteine substitution mutants give structural insight and identify ATP binding and activation sites at P2X receptors.
  63. (1997). Desensitization of the P2X(2) receptor controlled by alternative splicing.
  64. (1997). Different sensitivities to pH of ATP-induced currents at four cloned P2X receptors.
  65. (2001). Differential assembly of rat purinergic P2X7 receptor in immune cells of the brain and periphery.
  66. (1999). Differential modulation by copper and zinc of P2X2 and P2X4 receptor function.
  67. (1996). Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge.
  68. (1982). Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold.
  69. (1996). Domains of P2X receptors involved in desensitization.
  70. (1991). Effect of presynaptic P2 receptor stimulation on transmitter release.
  71. (1995). Electrophysiological properties of P2X-purinoceptors in rat superior cervical, nodose and guinea-pig coeliac neurones.
  72. (1993). Endothelin-1 enhances vasoconstrictor responses to exogenously administered and neurogenically released ATP in rabbit isolated perfused arteries.
  73. (2005). Ethanol differentially affects ATP-gated P2X3 and P2X4 receptor subtypes expressed in Xenopus oocytes.
  74. (2002). Ethanol sensitivity in ATP-gated P2X receptors is subunit dependent.
  75. (2007). Evidence for functional P2X4/P2X7 heteromeric receptors.
  76. (1990). Evidence for the formation of heteromultimeric potassium channels in Xenopus oocytes.
  77. (1992). Evidence that ATP released from the postsynaptic site by noradrenaline, is involved in mechanical responses of guinea-pig vas deferens: cascade transmission.
  78. (1993). Expression cloning of an ATP receptor from mouse neuroblastoma cells.
  79. (2000). Facilitation by P(2) receptor activation of acetylcholine release from rat motor nerve terminals: interaction with presynaptic nicotinic receptors.
  80. (2006). Fe65 interacts with P2X2 subunits at excitatory synapses and modulates receptor function.
  81. (1999). Functional and biochemical evidence for heteromeric ATP-gated channels composed of P2X1 and P2X5 subunits.
  82. (2004). Functional characterization of a P2X receptor from Schistosoma mansoni.
  83. (1998). Functional modulation of P2X2 receptors by cyclic AMP-dependent protein kinase.
  84. (2004). Functional regulation of P2X6 receptors by N-linked glycosylation: identification of a novel alpha beta-methylene ATP-sensitive phenotype.
  85. (2004). G-protein-coupled receptor regulation of P2X1 receptors does not involve direct channel phosphorylation.
  86. (2004). Gain and loss of channel function by alanine substitutions in the transmembrane segments of the rat ATP-gated P2X2 receptor.
  87. (2008). Gating the pore of P2X receptor channels.
  88. (1999). Hetero-oligomeric assembly of P2X receptor subunits. Specificities exist with regard to possible partners.
  89. (2004). Heterogeneity of P2X receptors in sympathetic neurons: contribution of neuronal P2X1 receptors revealed using knockout mice.
  90. (2002). Heteromerization of Kir2.x potassium channels contributes to the phenotype of Andersen's syndrome.
  91. (2002). Heteromultimeric P2X(1/2) receptors show a novel Pharmacol,
  92. (2001). Hsp90 ensures the transition from the early Ca2+-dependent to the late phosphorylation-dependent activation of the endothelial nitric-oxide synthase in vascular endothelial growth factor-exposed endothelial cells.
  93. (2002). Identification of a non-canonical tyrosine-based endocytic motif in an ionotropic receptor.
  94. (2001). Identification of a novel human ADP receptor coupled to G(i).
  95. (1998). Identification of a P2X1 purinoceptor expressed on human platelets.
  96. (2004). Identification of a trafficking motif involved in the stabilization and polarization of P2X receptors.
  97. (1994). Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the alpha subunit.
  98. (1998). Identification of amino acid residues contributing to desensitization of the P2X2 receptor channel.
  99. (1997). Identification of amino acid residues contributing to the pore of a P2X receptor.
  100. (2007). Identification of an intersubunit cross-link between substituted cysteine residues located in the putative ATP binding site of the P2X1 receptor.
  101. (2001). Identification of the platelet ADP receptor targeted by antithrombotic drugs.
  102. (1997). Immunohistochemical study of the P2X2 and P2X3 receptor subunits in rat and monkey sensory neurons and their central terminals.
  103. (2006). Impaired flow-dependent control of vascular tone and remodeling in P2X4-deficient mice.
  104. (2000). Inactivation of P2X2 purinoceptors by divalent cations.
  105. (2001). Inflammatory mediators potentiate ATP-gated channels through the P2X(3) subunit.
  106. (2000). Inhibition by ethanol of rat P2X(4) receptors expressed in Xenopus oocytes.
  107. (2003). Interaction between cysteines introduced into each transmembrane domain of the rat P2X2 receptor.
  108. (2001). International union of pharmacology. XXIV. Current status of the nomenclature and properties of P2X receptors and their subunits.
  109. (2003). Intracellular cross talk and physical interaction between two classes of neurotransmitter-gated channels.
  110. (1987). Involvement of calcium in calcium-current inactivation in smooth muscle cells from rat vas deferens.
  111. (1999). Ion permeation and block of P2X(2) purinoceptors: single channel recordings.
  112. (1996). Ionic permeability of, and divalent cation effects on, two ATP-gated cation channels (P2X receptors) expressed in mammalian cells.
  113. (2001). Ionic selectivity of native ATP-activated (P2X) receptor channels in dissociated neurones from rat parasympathetic ganglia.
  114. (1985). Is there a basis for distinguishing two types of P2-purinoceptor?
  115. (1995). K+ pore structure revealed by reporter cysteines at inner and outer surfaces.
  116. (2007). Lack of evidence for direct phosphorylation of recombinantly expressed P2X(2) and P2X (3) receptors by protein kinase C. Purinergic Signal,
  117. (1998). Membrane topology of an ATP-gated ion channel (P2X receptor).
  118. (2003). Modulation of 5-HT3 receptor-mediated response and trafficking by activation of protein kinase
  119. (2004). Modulation of human GABAA and glycine receptor currents by menthol and related monoterpenoids.
  120. (2002). Molecular physiology of P2X receptors.
  121. (2004). Molecular properties of ATP-gated P2X receptor ion channels.
  122. (2008). Molecular shape, architecture, and size of P2X4 receptors determined using fluorescence resonance energy transfer and electron microscopy.
  123. (2003). Monomeric and dimeric byproducts are the principal functional elements of higher order P2X1 concatamers.
  124. (2005). Mutagenesis studies of conserved proline residues of human P2X receptors for ATP indicate that proline 272 contributes to channel function.
  125. (2003). Mutation of a dibasic amino acid motif within the C terminus of the P2X7 nucleotide receptor results in trafficking defects and impaired function.
  126. (2002). Mutational analysis of the conserved cysteines of the rat P2X2 purinoceptor.
  127. (1999). Neighboring glycine residues are essential for P2X2 receptor/channel function.
  128. (1999). Neuronal P2X transmitter-gated cation channels change their ion selectivity in seconds.
  129. (1996). Neuronal synthesis, storage and release of ATP.
  130. (2001). Neurotransmitter/neuropeptide interactions in the regulation of neurohypophyseal hormone release.
  131. (1994). New structural motif for ligand-gated ion channels defined by an ionotropic ATP receptor.
  132. (1949). Nucleotides. Part II. A synthesis of adenosine triphosphate.
  133. (1929). On the phosohate fraction in muscle.
  134. (1878). On the Physiology of the Salivary Secretion: Part II. On the Mutual Antagonism of Atropin and Pilocarpin, having especial reference to their relations in the Sub-maxillary Gland of the Cat.
  135. (1996). On the use of thiol-modifying agents to determine channel topology.
  136. (1988). Origin of ATP release in the rat vas deferens: concomitant measurement of [3H]noradrenaline and [14C]ATP.
  137. (2008). Orthosteric and allosteric binding sites of P2X receptors.
  138. (1996). P2 purinoceptors: historical perspective and classification.
  139. (2000). P2X receptor expression in mouse urinary bladder and the requirement of P2X(1) receptors for functional P2X receptor responses in the mouse urinary bladder smooth muscle.
  140. (2004). P2X receptor subtype-specific modulation of excitatory and inhibitory synaptic inputs in the rat brainstem.
  141. (2002). P2X receptor trafficking in neurons is subunit specific.
  142. (2006). P2X receptors as cell-surface ATP sensors in health and disease.
  143. (1996). P2X receptors: a third major class of ligand-gated ion channels.
  144. (2002). P2X(1) receptor subunit contribution to gating revealed by a dominant negative PKC mutant.
  145. (2002). P2X(1) receptor-deficient mice establish the native P2X receptor and a P2Y6-like receptor in arteries.
  146. (1998). P2X1 and P2X3 receptors form stable trimers: a novel structural motif of ligand-gated ion channels.
  147. (2003). P2X1 receptor currents after disruption of the PKC site and its surroundings by dominant negative mutations in HEK293 cells.
  148. (1996). P2X4: an ATP-activated ionotropic receptor cloned from rat brain.
  149. (2007). Pannexin-1 couples to maitotoxin- and nigericin-induced interleukin-1beta release through a dye uptake-independent pathway.
  150. (2006). Parkin potentiates ATP-induced currents due to activation of P2X receptors in PC12 cells.
  151. (2004). patS minigenes inhibit heterocyst development of Anabaena sp. strain PCC 7120.
  152. (1995). Pharmacological characterization of heterologously expressed ATP-gated cation channels (P2x purinoceptors).
  153. (2005). Pharmacological profiles of cloned mammalian P2Y-receptor subtypes.
  154. (1999). Pore dilation of neuronal P2X receptor channels.
  155. (1995). Potassium channel assembly from concatenated subunits: effects of proline substitutions in S4 segments.
  156. (2005). Potentiation of P2X1 ATP-gated currents by 5-hydroxytryptamine 2A receptors involves diacylglycerol-dependent kinases and intracellular calcium.
  157. (1997). Presence of P2X1 purinoceptors in human platelets and megakaryoblastic cell lines.
  158. (1997). Primary structure and expression of a naturally truncated human P2X ATP receptor subunit from brain and immune system.
  159. (2005). Principles of pharmacology: the pathophysiologic basis of drug therapy.
  160. (2005). Profiling at recombinant homomeric and heteromeric rat P2X receptors identifies the suramin analogue NF449 as a highly potent P2X1 receptor antagonist.
  161. (2000). Properties of P2X and P2Y receptors are dependent on artery diameter in the rat mesenteric bed.
  162. (1996). Purinergic receptors: their role in nociception and primary afferent neurotransmission.
  163. (2001). Purinergic signalling: ATP release.
  164. (1994). Purinoceptors: are there families of P2X and P2Y purinoceptors?
  165. (1998). Receptors for purines and pyrimidines.
  166. (2000). Reduced vas deferens contraction and male infertility in mice lacking P2X1 receptors.
  167. (2009). Regions of the amino terminus of the P2X receptor required for modification by phorbol ester and mGluR1alpha receptors.
  168. (2000). Roles of individual N-glycans for ATP potency and expression of the rat P2X1 receptor.
  169. (1993). Selective inhibition of protein kinase C isozymes by the indolocarbazole Go 6976.
  170. (1999). Selectivity of diadenosine polyphosphates for rat P2X receptor subunits.
  171. (2002). Sensitization by extracellular Ca(2+) of rat P2X(5) receptor and its pharmacological properties compared with rat P2X(1).
  172. (1995). Site-directed mutagenesis of P2U purinoceptors. Positively charged amino acids in transmembrane helices 6 and 7 affect agonist potency and specificity.
  173. (1994). Structural determinant for assembly of mammalian K+ channels.
  174. (2000). Structural differences in the two agonist binding sites of the Torpedo nicotinic acetylcholine receptor revealed by time-resolved fluorescence spectroscopy.
  175. (1996). Structure of the NMDA receptor channel M2 segment inferred from the accessibility of substituted cysteines.
  176. (1997). Studies on the release and extracellular metabolism of endogenous ATP in rat superior cervical ganglion: support for neurotransmitter role of ATP.
  177. (2003). Subunit arrangement in P2X receptors.
  178. (2001). Subunit arrangement of gamma-aminobutyric acid type A receptors.
  179. (1996). Subunit positional effects revealed by novel heteromeric inwardly rectifying K+ channels.
  180. (1992). Subunit stoichiometry of a mammalian K+ channel determined by construction of multimeric cDNAs.
  181. (1999). Synaptic corelease of ATP and GABA in cultured spinal neurons.
  182. (1995). Synaptic desensitization of NMDA receptors by calcineurin.
  183. (1998). The 90-kDa molecular chaperone family: structure, function, and clinical applications. A comprehensive review.
  184. (1954). The capillary dilator substances in dry powders of spinal roots; a possible role of adenosine triphosphate in chemical transmission from nerve endings.
  185. (1996). The cytolytic P2Z receptor for extracellular ATP identified as a P2X receptor (P2X7).
  186. (1996). The exon sequence TAGG can inhibit splicing.
  187. (2001). The first transmembrane domain of the P2X receptor subunit participates in the agonist-induced gating of the channel.
  188. (1998). The heterotetrameric architecture of the epithelial sodium channel (ENaC).
  189. (1989). The human double-stranded DNA-activated protein kinase phosphorylates the 90-kDa heat-shock protein, hsp90 alpha at two NH2-terminal threonine residues.
  190. (2002). The intracellular tyrosine residues of the ATP-gated P2X(1) ion channel are essential for its function.
  191. (1959). The liberation of adenosine triphosphate on antidromic stimulation of sensory nerves.
  192. (2004). The nucleotide-binding domain of the Zn2+-transporting P-type ATPase from Escherichia coli carries a glycine motif that may be involved in binding of ATP.
  193. (1929). The physiological activity of adenine compounds with especial reference to their action upon the mammalian heart.
  194. (2000). The PSIPRED protein structure prediction server.
  195. (2002). The role of histidine residues in modulation of the rat P2X(2) purinoceptor by zinc and pH.
  196. (2000). The role of positively charged amino acids in ATP recognition by human P2X(1) receptors.
  197. (2000). The roles of P(2X1)and P(2T AC)receptors in ADP-evoked calcium signalling in human platelets.
  198. (1995). The selective P2X purinoceptor agonist, beta,gamma-methylene-L-adenosine 5'-triphosphate, discriminates between smooth muscle and neuronal P2X purinoceptors.
  199. (1998). The tetrameric structure of a glutamate receptor channel.
  200. (1957). The utilization of phosphate bond energy for sodium extrusion from giant axons.
  201. (2007). Thr339-to-serine substitution in rat P2X2 receptor second transmembrane domain causes constitutive opening and indicates a gating role for Lys308.
  202. (1998). Topological analysis of the ATP-gated ionotropic [correction of ionotrophic] P2X2 receptor subunit.
  203. (2008). TRPV1 shows dynamic ionic selectivity during agonist stimulation.
  204. (2003). Tuning activation of the AMPA-sensitive GluR2 ion channel by genetic adjustment of agonist-induced conformational changes.
  205. (2000). Two critical cysteine residues implicated in disulfide bond formation and proper folding of Kir2.1.
  206. (1993). Tyrosine kinase-dependent suppression of a potassium channel by the G protein-coupled m1 muscarinic acetylcholine receptor.
  207. (2003). Tyrosine phosphorylation of HSP90 within the P2X7 receptor complex negatively regulates P2X7 receptors.
  208. (2000). Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X3-deficient mice.
  209. (2005). Visualization of the trimeric P2X2 receptor with a crown-capped extracellular domain.
  210. (2000). Warm-coding deficits and aberrant inflammatory pain in mice lacking P2X3 receptors.

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