Leucine-2-alanine enkephalin induced delta opioid receptors internalization expressed stably in CHO cells.

Zhongguo yao li xue bao = Acta pharmacologica Sinica Pub Date : 1999-06-01

C H Wang, D H Zhou, J Chen, Z Q Chi

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Abstract

Aim: To characterize the internalization of delta opioid receptors (DOR) stably expressed in Chinese hamster ovary (CHO) cells and the role of the C-terminal in this process.

Methods: Receptor membrane anchoring was shown by immunofluorescence microscopy. Receptor internalization was assessed by measuring the radioligand binding resistant to the acid-buffer wash.

Results: Originally, all the wild-type (CHO-W) and C-truncated (CHO-T) DOR expressed were localized to the membrane. Agonist [3H] leucine-2-alanine enkephalin (LAE) but not the antagonist [3H]diprenorphine (Dip) induced rapid receptor internalization. The internalization of C-truncated DOR in CHO-T was similar to that of the wild-type in maximal level, but climbed up more slowly. DOR internalization was extracellular osmolarity- and temperature-sensitive. Pertussis toxin and universal protein kinase inhibitor staurosporine had no effect on it.

Conclusion: DOR internalization is an agonist and clathrin-coated pits dependent, but post-receptor cellular signal transduction independent process; moreover, the C-terminal of DOR, not engaged in membrane anchoring, affects the initialization of DOR internalization.

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亮氨酸-2-丙氨酸脑啡肽诱导δ阿片受体内化在CHO细胞中的稳定表达。

目的:研究δ阿片受体(DOR)在中国仓鼠卵巢(CHO)细胞中稳定表达的内化过程及其c端在这一过程中的作用。方法:采用免疫荧光显微镜观察受体膜锚定作用。受体内化是通过测量抗酸缓冲洗涤的放射性配体结合来评估的。结果:最初，所有野生型(CHO-W)和c -截断型(CHO-T) DOR的表达都局限于膜上。激动剂[3H]亮氨酸-2-丙氨酸脑啡肽(LAE)而非拮抗剂[3H]二丙诺啡(Dip)诱导受体快速内化。CHO-T中c -截断DOR的内在化在最大水平上与野生型相似，但上升速度较慢。DOR内化对细胞外渗透压和温度敏感。百日咳毒素和通用蛋白激酶抑制剂staurosporine对其无影响。结论:DOR内化是一个依赖于激动剂和网格蛋白包被凹坑的过程，但与受体后细胞信号转导无关;DOR的c端不参与膜锚定，影响DOR内化的初始化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Zhongguo yao li xue bao = Acta pharmacologica Sinica

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