Expression of dopamine D-1 receptor in Sf9 insect cells and agonism of l-12-chloroscoulerine on recombinant D-1 receptor

CORC > 上海药物研究所 > 中国科学院上海药物研究所 > 药理学第一研究室

	Expression of dopamine D-1 receptor in Sf9 insect cells and agonism of l-12-chloroscoulerine on recombinant D-1 receptor
	He, Y ; Jin, WQ ; Shen, QX ; Chen, XJ ; Jin, GZ
刊名	ACTA PHARMACOLOGICA SINICA
	2003-03
卷号	24 期号:3 页码:225-229
关键词	recombinant proteins dopamine D-1 receptors insects baculoviridae l-12-chloroscoulerine cyclic AMP
ISSN号	1671-4083
文献子类	Article
英文摘要	AIM: To express dopamine D, receptor in baculovirus-Sf9 cell system, and to investigate the effects of l-12-chloroscoulerine (l-CSL) on the recombinant D, receptor (D,R). METHODS: The recombinant baculovirus, Autographa californica nuclear polyhedrosis virus bearing D1R (AcNPV- D1R) was generated, and then was used to produce recombinant D1R in Sf9 insect cells. Expression of D1R in Sf9 cells was monitored by [H-3]SCH23390 binding assay. The effects of l-CSL on recombinant D,R were investigated by [H-3]SCH23390 binding assay and cAMP assay. RESULTS: The recombinant baculovirus AcNPV bearing D,R cDNA was generated, and was successfully expressed in Sf9 insect cells. The expression level of (B-max) was (0.94+/-0.06) nmol/g protein. The K-d value of [H-3]SCH23390 was (1.9+/-0.3) nmol/L, which was consistent with the previous results from calf striutam tissues. l-CSL had a high affinity to recombinant D1R with K-i value of (6.3+/-1.4) nmol/L, and increased the intracellular cAMP level in a concentration-dependent manner with EC50 value of 0.72 mumol/L and 95 % confidence limit was 0.67-0.77 mumol/L. Thus l-CSL has the D-1 receptor agonism. CONCLUSION: An efficient baculovirus-Sf9 insect cell system for dopamine D-1 receptor was constructed and l-CSL presented the D-1 receptor agonism on cellular-molecular level directly.
WOS关键词	12-CHLOROSCOULERINE ENANTIOMERS ; (+/-)12-CHLOROSCOULERINE ; (-)-STEPHOLIDINE ; BEHAVIORS ; NEURONS
WOS研究方向	Chemistry ; Pharmacology & Pharmacy
语种	英语
出版者	ACTA PHARMACOLOGICA SINICA
WOS记录号	WOS:000181439200006
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/274254]
专题	药理学第一研究室
通讯作者	Jin, WQ
作者单位	1.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Mat Med, Shanghai 200031, Peoples R China 2.Shanghai Inst Planned Parenthood Res, Shanghai 200032, Peoples R China
推荐引用方式 GB/T 7714	He, Y,Jin, WQ,Shen, QX,et al. Expression of dopamine D-1 receptor in Sf9 insect cells and agonism of l-12-chloroscoulerine on recombinant D-1 receptor[J]. ACTA PHARMACOLOGICA SINICA,2003,24(3):225-229.
APA	He, Y,Jin, WQ,Shen, QX,Chen, XJ,&Jin, GZ.(2003).Expression of dopamine D-1 receptor in Sf9 insect cells and agonism of l-12-chloroscoulerine on recombinant D-1 receptor.ACTA PHARMACOLOGICA SINICA,24(3),225-229.
MLA	He, Y,et al."Expression of dopamine D-1 receptor in Sf9 insect cells and agonism of l-12-chloroscoulerine on recombinant D-1 receptor".ACTA PHARMACOLOGICA SINICA 24.3(2003):225-229.