In vitro binding and signaling profile of the novel μ opioid receptor agonist endomorphin 2 in rat brain membranes

Mariana Spetea, Krisztina Monory, C. Tömböly, Géza Tóth, Eleni Tzavara, S. Benyhe, Jacques Hanoune, A. Borsodi

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Abstract

The recently discovered endogenous μ receptor selective endomorphin 2 was prepared in tritiated form by a catalytic dehalogenation method resulting in a specific radioactivity of 1.98 TBq/mmol (53.4 Ci/mmol), and used for in vitro labelling of rat brain membranes. The binding was saturable, stereospecific and of high affinity (K(d): 0.97 and 1.12 nM based on kinetic and equilibrium binding studies, respectively). The maximal number of binding sites (B(max)) was found to be 114.8 fmol/mg protein. [3H]Endomorphin 2 was displaced by μ-receptor selective specific peptides and heterocyclic compounds with high affinity, whereas κ and δ receptor specific ligands were much less potent. The K(i) values of endomorphin 1 and 2 in inhibiting [3H]naloxone binding increased by 15-fold in the presence of 100 mM NaCl which indicates the agonist property of these peptides. Endomorphins stimulated [35S]GTPγS binding and inhibited adenylyl cyclase activity which also provides evidence for the agonist character of endomorphins.

Original languageEnglish
Pages (from-to)720-725
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume250
Issue number3
DOIs
Publication statusPublished - Sep 29 1998

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Opioid Receptors
Rats
Brain
Membranes
Dehalogenation
Heterocyclic Compounds
Peptides
Radioactivity
Naloxone
Adenylyl Cyclases
Labeling
Binding Sites
Ligands
Kinetics
endomorphin 2
In Vitro Techniques
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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In vitro binding and signaling profile of the novel μ opioid receptor agonist endomorphin 2 in rat brain membranes. / Spetea, Mariana; Monory, Krisztina; Tömböly, C.; Tóth, Géza; Tzavara, Eleni; Benyhe, S.; Hanoune, Jacques; Borsodi, A.

In: Biochemical and Biophysical Research Communications, Vol. 250, No. 3, 29.09.1998, p. 720-725.

Research output: Contribution to journalArticle

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