Regular articles - Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins | |
Wei, Q; Zhou, DH; Shen, QX; Chen, J![]() ![]() | |
刊名 | CELL RESEARCH
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2000-06 | |
卷号 | 10期号:2页码:93-102 |
关键词 | human mu-opioid receptor (H mu OR) Sf9 insect cells pertussis toxin (PTX) endogenous G(i/o) proteins |
ISSN号 | 1001-0602 |
DOI | 10.1038/sj.cr.7290039 |
文献子类 | Article |
英文摘要 | Human mu-opioid receptor (H mu OR) with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells. The maximal binding capacity for the [H-3] diprenorphine and [H-3]ohmefentalnyl (Ohm) were 9.1 +/- 0.7 and 6.52 +/- 0.23 nmol/g protein, respectively. The [H-3] diprenorphine or [H-3] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by mu-selective agonists [D-ALa(2), N-methyl-Phe(4), glyol(5)] enkephalin (DAGO), Ohm, and morphine, but neither by delta nor by kappa selective agonist. Na+ (100 mM) and GTP (50 mu M) could reduce H mu OR agonists etorphine and Ohm affinity binding to the overexpressed H mu OR. mu-selective agonists DAGO and Ohm effectively stimulated [S-35] GTP gamma S binding (EC50 = 2.7 nM and 6.9 nM) and inhibited forskolin- stimulated cAMP accumulation (IC50 = 0.9 nM and 0.3 nM). The agonist-dependent effects could be blocked by opioid antagonist naloxone or by pretreatment of cells with pertussis toxin (PTX). These results demonstrated that H mu OR overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins. |
WOS关键词 | OPIATE-RECEPTOR ; HOMOLOGOUS DESENSITIZATION ; BACULOVIRUS EXPRESSION ; AGONIST ; ACTIVATION |
WOS研究方向 | Cell Biology |
语种 | 英语 |
CSCD记录号 | CSCD:805716 |
出版者 | SCIENCE PRESS |
WOS记录号 | WOS:000088017100002 |
内容类型 | 期刊论文 |
源URL | [http://119.78.100.183/handle/2S10ELR8/274617] ![]() |
专题 | 中国科学院上海药物研究所 |
通讯作者 | Zhou, DH |
作者单位 | 1.Chinese Acad Sci, Shanghai Acad Life Sci, Shanghai Inst Cell Biol, Shanghai 200031, Peoples R China 2.Chinese Acad Sci, Shanghai Acad Life Sci, Shanghai Inst Mat Med, Shanghai 200031, Peoples R China |
推荐引用方式 GB/T 7714 | Wei, Q,Zhou, DH,Shen, QX,et al. Regular articles - Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins[J]. CELL RESEARCH,2000,10(2):93-102. |
APA | Wei, Q.,Zhou, DH.,Shen, QX.,Chen, J.,Chen, LW.,...&Chi, ZQ.(2000).Regular articles - Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins.CELL RESEARCH,10(2),93-102. |
MLA | Wei, Q,et al."Regular articles - Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins".CELL RESEARCH 10.2(2000):93-102. |
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