Specific phospholipid requirements for the solubilization and reconstitution of D-1 dopamine receptors from striatal membranes.
Specific phospholipid requirements for the solubilization and reconstitution of D-1 dopamine receptors from striatal membranes.
复制标题
纹状体膜 D-1 多巴胺受体的溶解和重建需要特定的磷脂。
DOI：
10.1021/bi00172a034
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Sidhu,A
中科院分区：
文献类型：
--
作者：
Balen,P;Kimura,K;Sidhu,A
Revised Manuscript Received October 21, 1993s abstract: We have reported the solubilization and reconstitution of functional Dl dopamine receptors from rat striatal tissue, using sodium cholate as detergent [Sidhu, A.(1988) Biochemistry 27, 8768-8776]. Critical to our method of extraction was the absolute requirementfor the persistent presence of a crude extract of phospholipids (PLs) from bovine brain, during both solubilization of membranes and reconstitution of the soluble extract into PL vesicles. In the absence of PLs, fewer than 10% of the receptors were recovered, while in the presence of PLs, 40% of the receptors were reconstituted into vesicles. To probe the composition of PLs required by Dl dopamine receptors during these extraction procedures, specific PLs of a defined composition were used during either solubilization or reconstitution alone or during both solubilization and reconstitution. Phosphatidylcholine (PC), when used during the solublization procedure alone or during both solubilization and reconstitution, resulted in recovery of 41-48% of the Dl dopamine receptors but was 3.7-fold less effective when present during reconstitution alone (11%). Phosphatidylethanolamine (PE), when used during reconstitution alone, resulted in recovery of nearly 25% of the Dl dopamine receptors. When PE was present during either solubilization or both solubilization and reconstitution, 6-11% of the receptors were recovered. If PE was used with PC in ratios of 1: 1 or 2: 1, respectively, 28-38% of the receptors were recovered. When PL vesicles of PE: PC were present in ratios of 1: 2 during both solubilization and reconstitution, the maximum theoretical (74-87%) recovery of total receptor binding sites was achieved. These reconstituted receptors were pharmacologically active, with 53.5% of the receptor population present in the high-affinity state.Neuronal dopaminergic transmissions have been implicated in several neuropathological diseases including Parkinsonism, depression, drugaddiction, schizophrenia, hyperprolactemia, hypertension, and possibly Huntington’s chorea. Dopamine exerts its effects via two receptor subtypes, D-1 and D-2, which have been classified according to their ability to either stimulate or inhibit adenylate cyclase activation, respectively (Kebabian & Caine, 1979). Dl dopamine receptors have also been linked to diverse signaling pathways other than the adenylate cyclase
