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Effect of Guanine Nucleotides on [3H]Glutamate Binding and on Adenylate Cyclase Activity in Rat Brain Membranes

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Abstract

GMP-PNP, a non-hydrolyzable analog of GTP binds tightly to G-protein in the presence of Mg2+, so that the binding is stable even after exhaustive washings. This property was exploited to prepare membrane samples of rat brain where G-protein GTP-binding sites were saturated with GMP-PNP. Experiments carried out with these membranes showed that GTP, GMP-PNP, GDP-S and GMP (1 mM) inhibit the sodium-independent [3H]glutamate binding by 30–40% [F(4,40) = 5.9; p < .001], whereas only GMP-PNP activates adenylate cyclase activity [F(6,42) = 3.56; p < .01]. The inhibition of sodium-independent [3H]glutamate binding occurred in the absence of Mg2+. These findings suggest that guanine nucleotides may inhibit glutamate binding and activate adenylate cyclase through distinct mechanisms by acting on different sites.

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Authors and Affiliations

  1. Departamento de Quimica, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, 97119-900, Santa Maria, RS, Brasil

    Maribel A. Rubin

  2. Departamento de Bioquimica, Instituto de Biociências, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil

    Alan C. Medeiros, Paula C. B. Rocha, Carolina B. Livi & Diogo O. Souza

  3. Centro de Biologia Molecular (CSIC-UAM), Universidade Autónoma, Canto Blanco, 28049, Madrid, Spain

    Galo Ramirez

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  1. Maribel A. Rubin
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  6. Diogo O. Souza
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Rubin, M.A., Medeiros, A.C., Rocha, P.C.B. et al. Effect of Guanine Nucleotides on [3H]Glutamate Binding and on Adenylate Cyclase Activity in Rat Brain Membranes. Neurochem Res 22, 181–187 (1997). https://doi.org/10.1023/A:1027367624250

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