title: Method for isolation of kappa-opioid binding sites by dynorphin affinity chromatography.
abstract: A kappa-opioid receptor subtype was purified from a digitonin extract of frog brain membranes, using affinity chromatography. The affinity resin was prepared by coupling dynorphin (1-10) to AH Sepharose 4B. The purified receptor binds 4,750 pmol [3H]ethylketocyclazocine (EKC) per mg protein (5,600-fold purification over the membrane-bound receptor) with a Kd of 9.1 nM. The addition of cholesterol-phosphatidylethanolamine (2:1) enhanced 3.6-fold the binding activity of the purified material, which gives a purification very close to the theoretical. The purified receptor protein exhibits high affinity for kappa-selective ligands. The purified fraction shows one major band (65,000 Mr) in sodium dodecyl sulfate (SDS) gel electrophoresis.
authors.affiliation: Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, Szeged.
authors.forename: J
authors.initials: J
authors.lastname: Simon
authors.forename: S
authors.initials: S
authors.lastname: Benyhe
authors.forename: J
authors.initials: J
authors.lastname: Hepp
authors.forename: E
authors.initials: E
authors.lastname: Varga
authors.forename: K
authors.initials: K
authors.lastname: Medzihradszky
authors.forename: A
authors.initials: A
authors.lastname: Borsodi
authors.forename: M
authors.initials: M
authors.lastname: Wollemann
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D004399:Dynorphins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D011893:Rana esculenta; D011957:Receptors, Opioid / Q000302:isolation & purification*; D017473:Receptors, Opioid, kappa; D013347:Subcellular Fractions
chemical_list: D010446:Peptide Fragments; D011957:Receptors, Opioid; D017473:Receptors, Opioid, kappa; D004399:Dynorphins; C039666:dynorphin amide (1-10)