A simple and efficient method for solubilization of the dopamine receptor

Characterization of High‐Affinity Dopamine D2 Receptors and Modulation of Affinity States by Guanine Nucleotides in Cholate‐Solubilized Bovine Striatal Preparations

“…The cholic acid/NaCl-solubilized dopamine Dz receptor preparations fulfilled the essential criteria of solubilization, including (a) no decrease in the specific binding of [3H]spiroperidol and [3H]NPA following ultracentrifugation of the solubilized preparations at 100,000 g for 3 h, (b) ultrafiltration of the solubilized preparations through 0.22 pm filters without any loss of specific binding activity, and (c) complete absence of lamellar membrane fragments under electron microscopy. These observations are in close agreement with the published reports on the solubilization of brain dopamine D2 receptors using this cholic acid-NaCI combination (Hall et al, 1983;Ramwani and Mishra, 1983;Wouters et al, 1984).…”

“…Solubilization of dopamine D2 receptors coupled to the inhibitory component of guanine nucleotide regulatory proteins (G-proteins) is an essential step toward the complete understanding of the molecular mechanism of receptor-effector interactions. Recently, there have been several attempts to solubilize dopamine D2 receptors from different membrane preparations using various detergents (Madras et al, 1982;Hall et al, 1983;Kuno et al, 1983;Ramwani and Mishra, 1983;Lew and Goldstein, 1984;Wouters et al, 1984). In these studies, the kinetic and equilibrium binding parameters of the antagonist ([3H]spiroperidol) binding to the solubilized receptors are reported to be in close proximity to those for the native membrane-bound receptors.…”