Regular articles - Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins

doi:10.1038/sj.cr.7290039

	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; Chen, LW ; Wang, TL ; Pei, G ; Chi, ZQ
刊名	CELL RESEARCH
	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.