{"title":"免疫球蛋白 G1 Fc 片段标记的人阿片受体样受体保留了抑制 cAMP 积累的能力。","authors":"L Y Yung, K W Tsim, G Pei, Y H Wong","doi":"10.1159/000014645","DOIUrl":null,"url":null,"abstract":"<p><p>The human opioid receptor-like (ORL(1)) receptor was tagged with the immunoglobulin G1 Fc fragment at the carboxy-terminus and expressed in human embryonic kidney 293 cells. Expression of the ORL(1)-Fc receptor was confirmed by immunohistochemical staining. The fusion protein was enriched by affinity chromatography and then verified by immunodetection. The function of the ORL(1)-Fc receptor was determined by examining nociceptin/OFQ-induced inhibition of cAMP accumulation. The ORL(1)-Fc receptor inhibited the forskolin-stimulated cAMP accumulation. The inhibitory response was selectively induced by nociceptin/OFQ in a dose-dependent and pertussis toxin-sensitive manner. Our results indicate that the carboxy-terminal Fc-tagged ORL(1) receptor retained the ability to interact with G(i) proteins to inhibit adenylyl cyclase.</p>","PeriodicalId":79565,"journal":{"name":"Biological signals and receptors","volume":"9 5","pages":"240-7"},"PeriodicalIF":0.0000,"publicationDate":"2000-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000014645","citationCount":"0","resultStr":"{\"title\":\"Immunoglobulin G1 Fc fragment-tagged human opioid receptor-like receptor retains the ability to inhibit cAMP accumulation.\",\"authors\":\"L Y Yung, K W Tsim, G Pei, Y H Wong\",\"doi\":\"10.1159/000014645\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The human opioid receptor-like (ORL(1)) receptor was tagged with the immunoglobulin G1 Fc fragment at the carboxy-terminus and expressed in human embryonic kidney 293 cells. Expression of the ORL(1)-Fc receptor was confirmed by immunohistochemical staining. The fusion protein was enriched by affinity chromatography and then verified by immunodetection. The function of the ORL(1)-Fc receptor was determined by examining nociceptin/OFQ-induced inhibition of cAMP accumulation. The ORL(1)-Fc receptor inhibited the forskolin-stimulated cAMP accumulation. The inhibitory response was selectively induced by nociceptin/OFQ in a dose-dependent and pertussis toxin-sensitive manner. Our results indicate that the carboxy-terminal Fc-tagged ORL(1) receptor retained the ability to interact with G(i) proteins to inhibit adenylyl cyclase.</p>\",\"PeriodicalId\":79565,\"journal\":{\"name\":\"Biological signals and receptors\",\"volume\":\"9 5\",\"pages\":\"240-7\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2000-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1159/000014645\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biological signals and receptors\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1159/000014645\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biological signals and receptors","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1159/000014645","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
人阿片受体样（ORL(1)）受体在羧基末端被免疫球蛋白 G1 Fc 片段标记，并在人胚肾 293 细胞中表达。免疫组织化学染色证实了 ORL(1)-Fc 受体的表达。融合蛋白通过亲和层析富集，然后通过免疫检测进行验证。通过检测痛觉素/OFQ 诱导的 cAMP 积累抑制作用，确定了 ORL(1)-Fc 受体的功能。ORL(1)-Fc 受体抑制了福斯可林刺激的 cAMP 积累。痛觉素/OFQ以剂量依赖和百日咳毒素敏感的方式选择性地诱导抑制反应。我们的研究结果表明，羧基末端 Fc 标记的 ORL（1）受体保留了与 G（i）蛋白相互作用抑制腺苷酸环化酶的能力。
Immunoglobulin G1 Fc fragment-tagged human opioid receptor-like receptor retains the ability to inhibit cAMP accumulation.
The human opioid receptor-like (ORL(1)) receptor was tagged with the immunoglobulin G1 Fc fragment at the carboxy-terminus and expressed in human embryonic kidney 293 cells. Expression of the ORL(1)-Fc receptor was confirmed by immunohistochemical staining. The fusion protein was enriched by affinity chromatography and then verified by immunodetection. The function of the ORL(1)-Fc receptor was determined by examining nociceptin/OFQ-induced inhibition of cAMP accumulation. The ORL(1)-Fc receptor inhibited the forskolin-stimulated cAMP accumulation. The inhibitory response was selectively induced by nociceptin/OFQ in a dose-dependent and pertussis toxin-sensitive manner. Our results indicate that the carboxy-terminal Fc-tagged ORL(1) receptor retained the ability to interact with G(i) proteins to inhibit adenylyl cyclase.
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