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The structure of antagonist-bound m1-LBR is presented. Also, analysis of the Gd-binding site revealed details of the LB2-LB2 contact site, which consists of four acidic residues. Out of the four acidic residues, Glu 238 is conserved among all subtypes and Asp 242 is replaced by glutamate in other subtypes. Furthermore, Asp 191 and Glu 231 are found in the surrounding area, and thus this site has a high negative charge.
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3+. Proc. Natl. Acad. Sci. U.S.A. 99:2002;2660-2665 The structure of antagonist-bound m1-LBR is presented. Also, analysis of the Gd-binding site revealed details of the LB2-LB2 contact site, which consists of four acidic residues. Out of the four acidic residues, Glu 238 is conserved among all subtypes and Asp 242 is replaced by glutamate in other subtypes. Furthermore, Asp 191 and Glu 231 are found in the surrounding area, and thus this site has a high negative charge.
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Mutation analyses of mGluR1 ligand binding residues are presented. The magnitude of the ligand-binding capacities and the amplitude of the intracellular signaling were almost correlated. Alanine substitution of the four residues that interact with the α-amino group of glutamate in the crystal abolished responses both to glutamate and quisqualate. Mutations in the three residues that interact with the γ-carboxy group of glutamate caused a loss of responsiveness to glutamate but not to quisqualate. The main five residues in the LB1 dimer interface were also mutated; a mutant in which Ile was replaced with Ala at position 120 lost the capacity for signal propagation.
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Molecular determinants of high affinity binding to Group III metabotropic glutamate receptors
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3H ]L-AP4) binding to the membrane preparations was examined. The authors discuss the binding mode of L-serine-O-phosphate, a potential endogenous compound, to an m4-LBR model.
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Identification of essential residues involved in the glutamate-binding pocket of the Group II metabotropic glutamate receptor
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35S ]GTPγS binding and activation of GIRK current were examined.
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D'Souza-Li L., Yang B., Canaff L., Bai M., Hanley D.A., Bastepe M., Salisbury S.R., Brown E.M., Cole D.E.C., Hendy G.N. Identification and functional characterization of novel calcium-sensing receptor mutations in familial hypocalciuric hypercalcemia and autosomal dominant hypocalcemia. J. Clin. Endocrinol. Metab. 87:2002;1309-1318.
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25
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Tas1r3, encoding a new candidate taste receptor, is allelic to the sweet responsiveness locus Sac
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The mutated residue in a mouse strain in which the sense of taste was abolished was investigated. A mutation whereby Ala replaces Ile at position 160 in T1r3 produces an extra glycosylation site at the residue corresponding to that of the mGluR1 LB1 dimer interface. The authors speculate that this would interfere with dimerization.
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Max M., Shanker Y.G., Huang L., Rong M., Liu Z., Campagne F., Weinstein H., Damak S., Margolskee R.F. Tas1r3, encoding a new candidate taste receptor, is allelic to the sweet responsiveness locus Sac. Nat. Genet. 28:2001;58-63 The mutated residue in a mouse strain in which the sense of taste was abolished was investigated. A mutation whereby Ala replaces Ile at position 160 in T1r3 produces an extra glycosylation site at the residue corresponding to that of the mGluR1 LB1 dimer interface. The authors speculate that this would interfere with dimerization.
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Closure of the venus flytrap module of mGluR8 receptor and the activation process: Insights from mutations converting antagonists into agonists
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Various chimeric combinations of GB1 and GB2 subunits were constructed and their ligand binding properties and other functionalities, including G-protein coupling, were examined. The GB2 cytoplasmic region contained the determinant for G-protein coupling; however, the GB1 cytoplasmic region improved the coupling efficacy. The GB2 subunit increases GABA affinity to the GB1 subunit. These data reveal that there are multiple allosteric interactions between the two subunits.
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B receptor function. EMBO J. 20:2001;2152-2159 Various chimeric combinations of GB1 and GB2 subunits were constructed and their ligand binding properties and other functionalities, including G-protein coupling, were examined. The GB2 cytoplasmic region contained the determinant for G-protein coupling; however, the GB1 cytoplasmic region improved the coupling efficacy. The GB2 subunit increases GABA affinity to the GB1 subunit. These data reveal that there are multiple allosteric interactions between the two subunits.
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CaR and mGluR1α were co-immunoprecipitated from bovine thalamus. The CaR-mGluR1α heterodimer was detected when two receptors were heterologously expressed in human embryonic kidney (HEK)-293 cells. The heterodimer is sensitive to both agonists.
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Gama L., Wilt S.G., Breitwieser G.E. Heterodimerization of calcium sensing receptors with metabotropic glutamate receptors in neurons. J. Biol. Chem. 276:2001;39053-39059 CaR and mGluR1α were co-immunoprecipitated from bovine thalamus. The CaR-mGluR1α heterodimer was detected when two receptors were heterologously expressed in human embryonic kidney (HEK)-293 cells. The heterodimer is sensitive to both agonists.
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Gama, L.1
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0242460494
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Metabotropic glutamate 1α and adenosine A1 receptors assemble into functionally interacting complexes
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An interaction between mGluR1α and adenosine A1 receptor was revealed by co-immunoprecipitation of rat cerebellum synaptosome and co-transfected cells. In the cells, glutamate/adenosine synergism was observed. Cytoplasmic regions seemed to be involved in the interaction, as a subtype mGluR1β does not interact with the A1 receptor.
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Ciruela F., Escriche M., Buergueño J., Angulo E., Casadó V., Soloviev M.M., Canela E.I., Mallol J., Chan W.-Y., Lluis C.et al. Metabotropic glutamate 1α and adenosine A1 receptors assemble into functionally interacting complexes. J. Biol. Chem. 276:2001;18345-18351 An interaction between mGluR1α and adenosine A1 receptor was revealed by co-immunoprecipitation of rat cerebellum synaptosome and co-transfected cells. In the cells, glutamate/adenosine synergism was observed. Cytoplasmic regions seemed to be involved in the interaction, as a subtype mGluR1β does not interact with the A1 receptor.
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Ciruela, F.1
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34
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0036415131
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Probing intermolecular protein-protein interactions in the calcium-sensing receptor homodimer using bioluminescence resonance energy transfer (BRET)
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Pre-existing dimerization of CaR was probed by BRET techniques. The authors discuss causes of the lack of BRET signal change upon ligand challenge.
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Jensen A.A., Hansen J.L., Sheikh S.P., Bräuner-Osborne H. Probing intermolecular protein-protein interactions in the calcium-sensing receptor homodimer using bioluminescence resonance energy transfer (BRET). Eur. J. Biochem. 269:2002;5076-5087 Pre-existing dimerization of CaR was probed by BRET techniques. The authors discuss causes of the lack of BRET signal change upon ligand challenge.
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2+-sensing function of the metabotropic glutamate receptors. Science. 279:1998;1722-1725.
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Construction of a high affinity zinc binding site in the mtabotropic glutamate receptor mGLuR1. Noncompetitive antagonism originating from the amino-terminal domain of a family C G-protein-coupled receptor
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2+ binding prevents transition from the resting state to the activated state because of steric interaction at the LB2 contact site.
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0031766635
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An antagonist peptide-EPO receptor complex suggests that receptor dimerization is not sufficient for activation
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Livnah O., Johnson D.L., Stura E.A., Farrell F.X., Barbone F.P., You Y., Liu K.D., Goldsmith M.A., He W., Krause C.D.et al. An antagonist peptide-EPO receptor complex suggests that receptor dimerization is not sufficient for activation. Nat. Struct. Biol. 5:1998;993-1004.
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Livnah O., Stura E.A., Middleton S.A., Johnson D.L., Jolliffe L.K., Wilson I.A. Crystallographic evidence for preformed dimers of erythropoietin receptor before ligand activation. Science. 283:1999;987-990.
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41
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This paper contains further analysis of the findings of [41], which presented the first determination of the hormone-binding domain of NPR-A in the ligand-free state. The two protomers are connected at their C-terminal domains.
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•], the two protomers were connected at their N-terminal domains. The two C-terminal domains approached each other in the ligand-bound crystal structure. The stoichiometry of the ligand and receptor and the thermodynamics of the hormone-binding process were also analyzed, using calorimetry.
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Structural determination of the Ach-binding protein of snail revealed that the proteins associate as pentamers. Hepes buffer molecules are bound in the subunit interfaces, possibly at Ach-binding sites.
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A chemical compound, BAY36-7620, which shares no structural similarity with glutamate, inhibits the basal activity of mGluR1 when transiently expressed in HEK-293 cells. This inhibition of the constitutive activity defines the inverse agonism. Chimeric receptor experiments show the interaction of the compound with the transmembrane region of mGluR1.
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Two series of chemical compounds enhanced glutamate responses in native (from rat brain slices) and recombinant mGluR1. This allosteric potentiation site is postulated to be in the transmembrane region. The authors discuss the involvement of helix III in the transmembrane region.
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