RECEPTOR-MEDIATED INHIBITION OF ADENYLATE CYCLASE

这项研究计划的长期目标是深入了解 受体抑制腺苷环化酶活性的作用机制。 包括阿片类药物、D-2多巴胺能、 α2-肾上腺素能、嘌呤能和胆碱能被认为是调节 抑制腺苷环化酶的神经元活性。因为这些受体 在调节大脑功能、洞察力方面发挥关键作用 对其作用机制的了解是理解和 涉及这些受体的神经疾病的治疗。最近，一位 具有类似脑的药理特性的MU-阿片受体 在一种分泌催乳素的细胞上发现了MU-阿片受体 肿瘤命名为7315c。初步结果表明，这种受体是 与抑制GTP结合蛋白有关，称为Ni，它起作用 作为抑制性受体和腺苷环化酶之间的转导。这个 这项研究提案的具体目的是确定Mu-阿片剂如何 受体增强鸟苷与镍的相互作用。它是 假设Mu-阿片受体的激活增强了 尼泊尔国内生产总值换算成国内生产总值。最初，将对鸦片类药物进行测试 刺激7315c膜中GTP酶活性的能力。下一步，GDP将是 测试其阻止GPP(NH)p-和GTP诱导的 镍的活化。阿片激动剂也将接受测试，以确定它们是否有能力 加强从镍中去除GPP(NH)p并导致可预见的变化 腺苷环化酶活力。百日咳毒素最近被提议用来 诱导镍的ADP核糖基化，使镍和镍解偶联 抑制性受体。百日咳毒素将被测试其能力 阻断Mu-阿片受体介导的鸟苷核苷酸交换。它 也将确定百日咳毒素是否对 抑制性受体。含有抑制性物质的中间叶膜 D-2多巴胺受体将与百日咳毒素处理后融合 7315c膜(不含D-2受体)。D-2多巴胺 然后将测试其与镍和镍重新偶联的能力 抑制腺苷环化酶。最后，将使用阿片剂亲和柱 试图分离与Mu-阿片受体密切相关的Mu-阿片受体 你好。

The long term objective of this research plan is to gain insight into the mechanism of action of receptors inhibiting adenylate cyclase activity. Numerous neuronal receptors including opiate, D-2 dopaminergic, alpha2-adrenergic, purinergic, and cholinergic are believed to modulate neuronal activity by inhibiting adenylate cyclase. Since these receptors play a critical role in modulating the functioning of the brain, insight into their mechanism of action is fundamental to the understanding and treatment of neurological disorders involving these receptors. Recently, a Mu-opiate receptor with pharmacological properties similar to brain Mu-opiate receptors was discovered on the cells of a prolactin secreting tumor termed 7315c. Preliminary results suggest that this receptor is associated with the inhibitory GTP binding protein, termed Ni, which acts as a transducer between the inhibitory receptor and adenylate cyclase. The specific aim of this research proposal is to determine how the Mu-opiate receptor enhances the interaction of guanyl nucleotides with Ni. It is hypothesized that activation of the Mu-opiate receptor enhances the exchange of GDP for GTP at Ni. Initially, opiates will be tested for their ability to stimulate GTPase activity in 7315c membranes. Next, GDP will be tested for its ability to block both the Gpp(NH)p- and the GTP-induced activation of Ni. Opiate agonists will also be tested for their ability to enhance the removal of Gpp(NH)p from Ni and cause predictable changes in adenylate cyclase activity. Pertussis toxin has recently been proposed to induce an ADP ribosylation of Ni resulting in the uncoupling of Ni and inhibitory receptors. Pertussis toxin will be tested for its ability to block Mu-opiate receptor-mediated exchange of guanyl nucleotides at Ni. It will also be determined if pertussis toxin has a direct effect on inhibitory receptors. Intermediate lobe membranes containing an inhibitory D-2 dopamine receptor will be treated with pertussis toxin and then fused with 7315c membranes (which contain no D-2 receptor). The D-2 dopamine receptor will then be tested for its ability to recouple with Ni and inhibit adenylate cyclase. Finally, an opiate affinity column will be used in an attempt to isolate the Mu-opiate receptor in close association with Ni.