Affinity labeling of δ opioid receptors by an enkephalin-derivative alkylating agent, DSLET-Mal

Ildikó Szatmári, G. Orosz, Kálmán Medzihradszky, A. Borsodi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Opioid binding properties of Tyr-D-Ser-Gly-Phe-Leu-Thr-NH-NH-Gly-Mal (DSLET-Mal), a novel enkephalin-framed affinity label, was determined in rat brain membranes. In competition studies the ligand showed high affinity for the delta opioid sites, labelled by [[3H][Ile5,6]deltorphin II (K(i) = 8 nM), whereas its binding to the mu ([3H]DAMGO) and kappa ([3H]EK(C) sites was weaker. Preincubation of the rat brain membranes with DSLET-Mal at micromolar concentrations resulted in a wash-resistant and dose dependent inhibition of the [3H][Ile5,6]deltorphin II binding sites (96% blocking at 10 μM concentration). Intracerebroventricular (ICV) administration of DSLET-Mal reduced the density of delta opioid receptors and had no effect on mu and kappa receptors, as determined by saturation binding studies. [Ile5,6]deltorphin II-stimulated [35S]GTPγS binding was determined in membrane preparations of different brain areas of the ICV-treated animals. In both frontal cortex and hippocampus DSLET-Mal significantly decreased G protein activation by the delta agonist, having no effect on DAMGO stimulated [35S]GTPγS binding. DSLET-Mal had qualitatively similar effects on both receptor binding and G protein activation. These characteristics of the compound studied suggest that DSLET-Mal can serve as an affinity label for further studies of the delta-opioid receptors.

Original languageEnglish
Pages (from-to)513-519
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume265
Issue number2
DOIs
Publication statusPublished - Nov 19 1999

Fingerprint

Enkephalins
Alkylating Agents
Opioid Receptors
Affinity Labels
Ala(2)-MePhe(4)-Gly(5)-enkephalin
Labeling
Brain
delta Opioid Receptor
Derivatives
Membranes
GTP-Binding Proteins
Opioid Analgesics
Rats
Chemical activation
kappa Opioid Receptor
mu Opioid Receptor
Frontal Lobe
Hippocampus
Carrier Proteins
Animals

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Affinity labeling of δ opioid receptors by an enkephalin-derivative alkylating agent, DSLET-Mal. / Szatmári, Ildikó; Orosz, G.; Medzihradszky, Kálmán; Borsodi, A.

In: Biochemical and Biophysical Research Communications, Vol. 265, No. 2, 19.11.1999, p. 513-519.

Research output: Contribution to journalArticle

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