Separation of κ‐opioid receptor subtype from frog brain

FEBS Letters

Borsodi, Anna¹  Simon, József¹  Benyhe, Sándor¹  Wollemann, Mária¹  Szücs, Mária¹ 

[1] Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, 6701 Szeged, PO Box 521, Hungary

关键词: Opioid receptor subtype;    Solubilization;    Separation;    Sucrose density gradient centrifugation;    Gel filtration;    Hydrodynamical parameters;    EKC;    ethylketocyclazocine;    DAGO;    Tyr-D-Ala2-Me-Phe4-Gly-ol5-enkephalin;    DALA;    Tyr-D-Ala2-L-Leu5-enkephalin;    DADLE;    Tyr-D-Ala5-D-Leu5-enkephalin;    PMSF;    phenylmethylsulfonyl fluoride;    CHAPS;    3-[(3-cholamidopropyl)dimethylammonio]-1 -propanesulfonate;   

DOI  :  10.1016/0014-5793(85)80818-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Complete separation of the [³H]ethylketocyclazocine ([³H]EKC) specific binding (k subtype) from tritiated Tyr-D-Ala²-Me-Phe⁴-Gly-ol⁵ enkephalin (DAGO) and Tyr-D-Ala²-L-Leu⁵-enkephalin (DALA) binding (μ-and δ-subtypes, respectively) was achieved by Sepharose-6B chromatography and sucrose density gradient centrifugation of digitonin solubilized frog brain membranes. The apparent sedimentation coefficient (S ₂₀,w) for the k receptor-detergent complex was 13.1 S and the corresponding Stokes radius 64 Å. The isolated fractions exhibited high affinity for EKC and bremazocine, whereas μ- and δ-specific ligands were unable to compete for the [³H]EKC binding sites, indicating that the κ subtype represents a separate molecular entity from the μ and δ receptor sites.

【 授权许可】

Unknown