Effect of guanine nucleotides on [3H]glutamate binding and on adenylate cyclase activity in rat brain membranes

Neurochem Res. 1997 Feb;22(2):181-7. doi: 10.1023/a:1027367624250.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Animals
  • Brain / drug effects
  • Brain / metabolism*
  • Cell Membrane / metabolism
  • GTP-Binding Proteins / metabolism
  • Glutamic Acid / metabolism*
  • Guanine Nucleotides / metabolism
  • Guanine Nucleotides / pharmacology*
  • Guanosine Diphosphate / analogs & derivatives
  • Guanosine Diphosphate / pharmacology
  • Guanosine Monophosphate / pharmacology
  • Guanosine Triphosphate / metabolism
  • Guanylyl Imidodiphosphate / metabolism
  • Magnesium / pharmacology
  • Male
  • Rats
  • Rats, Wistar
  • Sodium / pharmacology
  • Thionucleotides / pharmacology
  • Tritium

Substances

  • Guanine Nucleotides
  • Thionucleotides
  • Tritium
  • Guanosine Diphosphate
  • Guanylyl Imidodiphosphate
  • Glutamic Acid
  • guanosine 5'-O-(2-thiodiphosphate)
  • Guanosine Monophosphate
  • Guanosine Triphosphate
  • Sodium
  • GTP-Binding Proteins
  • Adenylyl Cyclases
  • Magnesium