Purification of the opiate receptor of NG108-15 neuroblastoma-glioma hybrid cells.
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| Abstract |
Opiate receptors from NG108-15 neuroblastoma-glioma hybrid cell membranes were purified to apparent homogeneity in a form covalently labeled with the opiate affinity ligand 3-methylfentanylisothiocyanate (super-FIT). The purification procedure consists of five steps: quantitative labeling of opiate receptors in membranes with super-FIT; solubilization in a lubrol/3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate mixture; adsorption to and elution from a wheat germ agglutinin-Sepharose column; immunoaffinity chromatography on columns of immobilized anti-fentanyl; preparative gel electrophoresis in the presence of NaDodSO4. The protein was purified approximately 30,000-fold in 3% overall yield to a state in which there is 1 mol of super-FIT bound per mol of protein, corresponding to 21,000 pmol per mg of protein, the theoretically expected specific activity. The protein is glycosylated and migrates on NaDodSO4 gel electrophoresis with a Mr near 58,000. It has a strong tendency to dimerize, even in the presence of denaturing detergents, and it exists primarily as an oligomer in nondenaturing detergents.
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| Authors | W F Simonds, T R Burke Jr, K C Rice, A E Jacobson, W A Klee |
| Journal | Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 82 Issue 15 Pg. 4974-8 (Aug 1985) ISSN: 0027-8424 [Print] United States |
| PMID | 2991902 (Publication Type: Journal Article) |
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