Purification of a kappa-opioid receptor subtype from frog brain

J. Simon, S. Benyhe, J. Hepp, A. Khan, A. Borsodi, M. Szücs, K. Medzihradszky, M. Wollemann

Research output: Contribution to journalArticle

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Abstract

A kappa-opioid receptor subtype was purified from a digitonin solubilized preparation of frog brain membranes using affinity chromatography. The affinity resin was prepared by coupling D-Ala2-Leu3-enkephalin to Sepharose-6B matrix. After elution of the receptor by 50 μmol naloxone, the kappa-subtype was separated from the mu- and delta-subtypes by gel permeation chromatography on Sepharose-6B. The purified receptor binds 3,900 pmol [3H]-ethylketocyclazocine per mg protein (a 4,300-fold purification over the membrane-bound receptor) with a KD of 8.3 nM. The purified receptor protein exhibits high affinity for kappa-selective ligands. The purified fraction shows two bands (Mr 65,000 and 58,000) in sodium dodecyl sulfate gel electrophoresis.

Original languageEnglish
Pages (from-to)19-28
Number of pages10
JournalNeuropeptides
Volume10
Issue number1
DOIs
Publication statusPublished - 1987

Fingerprint

kappa Opioid Receptor
Anura
Sepharose
Purification
Brain
Ethylketocyclazocine
Membranes
Affinity chromatography
Digitonin
Enkephalins
Gel permeation chromatography
Naloxone
Electrophoresis
Affinity Chromatography
Sodium Dodecyl Sulfate
Gel Chromatography
Proteins
Resins
Gels
Ligands

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Endocrinology, Diabetes and Metabolism
  • Clinical Neurology
  • Neuroscience(all)
  • Cellular and Molecular Neuroscience

Cite this

Purification of a kappa-opioid receptor subtype from frog brain. / Simon, J.; Benyhe, S.; Hepp, J.; Khan, A.; Borsodi, A.; Szücs, M.; Medzihradszky, K.; Wollemann, M.

In: Neuropeptides, Vol. 10, No. 1, 1987, p. 19-28.

Research output: Contribution to journalArticle

Simon, J. ; Benyhe, S. ; Hepp, J. ; Khan, A. ; Borsodi, A. ; Szücs, M. ; Medzihradszky, K. ; Wollemann, M. / Purification of a kappa-opioid receptor subtype from frog brain. In: Neuropeptides. 1987 ; Vol. 10, No. 1. pp. 19-28.
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