Heterologous expression of lipoprotein-associated phospholipase A(2) in different expression systems

doi:10.1016/j.pep.2006.03.009

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	Heterologous expression of lipoprotein-associated phospholipase A(2) in different expression systems
	Zhang, Fujun ; Dong, Li ; Cai, Maojun ; Shen, Jingkang ; Wang, Yiping
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2006-08
卷号	48 期号:2 页码:300-306
关键词	lipoprotein-associated phospholipase A(2) atherosclerosis coronary baculovirus insect cells Pichia pastoris expression purification
ISSN号	1046-5928
DOI	10.1016/j.pep.2006.03.009
文献子类	Article
英文摘要	Lipoprotein-associated phospholipase A, (Lp-PLA(2)) is a key enzyme involved in atherosclerosis, and has been considered as a new target for drug discovery. The major difficulty for high-throughput screening of Lp-PLA(2) inhibitors and for functional studies was their fast and efficient production. Purification of native Lp-PLA(2) from human plasma was complicated and produced a very low yield. We herein examined the feasibility of expressing and purifying recombinant Lp-PLA(2) in different heterologous expression systems. The fusion Lp-PLA(2) was expressed at high levels and exhibited strong enzyme activity in insect cell-baculovirus expression system. The functional enzyme could also be produced in Pichia pastoris. The inclusion of a Kozak sequence increased greatly the expression level of recombinant Lp-PLA(2) in insect cells, but had little effect on the expression of recombinant Lp-PLA(2) in P. pastoris and Escherichia coli. P. pastoris-produced Lp-PLA(2) could be purified rapidly and conveniently through a one-step procedure, while baculovirus-produced Lp-PLA(2) could be efficiently purified through a two-step procedure. This ability to readily produce recombinant Lp-PLA(2) could provide a screening model for Lp-PLA(2) inhibitors and will facilitate further studies on this enzyme. (c) 2006 Elsevier Inc. All rights reserved.
WOS关键词	ACTIVATING-FACTOR-ACETYLHYDROLASE ; CORONARY-HEART-DISEASE ; ORALLY-ACTIVE INHIBITORS ; C-REACTIVE PROTEIN ; MIDDLE-AGED MEN ; THERAPEUTIC TARGET ; FOLLOW-UP ; RISK ; ATHEROSCLEROSIS ; POTENT
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000239696900015
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/273532]
专题	药理学第一研究室中科院受体结构与功能重点实验室新药研究国家重点实验室
通讯作者	Wang, Yiping
作者单位	1.Chinese Acad Sci, Shanghai Inst Biol Studies, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 201203, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Biol Studies, Shanghai Inst Mat Med, Dept Med Chem, Shanghai 201203, Peoples R China 3.Chinese Acad Sci, Grad Sch, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Fujun,Dong, Li,Cai, Maojun,et al. Heterologous expression of lipoprotein-associated phospholipase A(2) in different expression systems[J]. PROTEIN EXPRESSION AND PURIFICATION,2006,48(2):300-306.
APA	Zhang, Fujun,Dong, Li,Cai, Maojun,Shen, Jingkang,&Wang, Yiping.(2006).Heterologous expression of lipoprotein-associated phospholipase A(2) in different expression systems.PROTEIN EXPRESSION AND PURIFICATION,48(2),300-306.
MLA	Zhang, Fujun,et al."Heterologous expression of lipoprotein-associated phospholipase A(2) in different expression systems".PROTEIN EXPRESSION AND PURIFICATION 48.2(2006):300-306.