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  • Title: [3H]clonidine and [3H]yohimbine binding to solubilized alpha 2-adrenoceptors from rat cerebral cortex.
    Author: Kitamura Y, Tanaka H, Nomura Y.
    Journal: Eur J Pharmacol; 1986 Apr 16; 123(2):263-70. PubMed ID: 3011462.
    Abstract:
    Alpha 2-adrenoceptors were solubilized from rat cerebral cortex using the zwitterionic detergent, 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate (CHAPS). The CHAPS extract retained binding activity for [3H]clonidine and [3H]yohimbine. Treatment of membranes with 10 mM CHAPS solubilized about 30% of the [3H]clonidine binding sites in the starting membranes. A Scatchard plot of [3H]clonidine binding to the CHAPS extract showed a non-linear curve, indicating the existence of the two distinct binding components. The effects of GTP and cations on alpha 2-agonist and antagonist binding to the CHAPS extract were similar to the effects in membrane preparations. Sepharose CL-4B column chromatography showed the alpha 2-agonist binding complex to be a larger molecule, with a Stokes radius of 85 A, than the alpha 2-antagonist binding complex with a radius of 71 A. These results indicate that the complexes between the alpha 2-adrenoceptors and GTP binding regulatory proteins remain intact throughout the CHAPS solubilization procedure.
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