Purification and reconstitution of the δ opioid receptor

FEBS Letters

Gomathi, K.G.¹  Sharma, Shall K.¹ 

[1] Department of Biochemistry, All India Institute of Medical Sciences, New Delhi 110029, India

关键词: δ Opioid receptor;    NG108-15;    Reconstitution;    CHAPS;    3-[(3-cholamidopropyl)dimethyl ammonio]1-prppane-sulfonate;    DADLE;    d-Ala2;    d-Leu5-enkephalin;    DPDPE;    D-Pen2;    D-Pen5-enkephalin;    ELISA;    enzyme-linked immunosorbent assay;    IMS;    immune mouse serum;    Kd;    equilibrium dissociation constant;    kDa;    kilodaltons;    PIMS;    pre-immune mouse serum;    PMSF;    phenyl methyl sulfonyl fluoride;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    WGA;    wheat germ agglutinin;   

DOI  :  10.1016/0014-5793(93)80261-R

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The δ opioid receptor has been purified, in an active form, by succinylmorphine affinity chromatography. The receptor was purified partially from bovine frontal cortex and to apparent homogeneity from neuroblastoma × glioma hybrid NG108-15 cells as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining. Antiserum to the purified bovine receptor inhibited ligand binding to membranes and immunoprecipitated a 58 kDa protein from NG108-15 cells. Reconstitution of the receptor with lipids enhanced binding by 9-fold. The 58 kDa band protein after electroelution and reconstitution with lipids also showed specific binding, indicating that the receptor could be renatured even after SDS-PAGE in an appropriate lipid environment.

【 授权许可】

Unknown