Tailoring the pH Dependence of Human Non-pancreatic Secretory   Phospholipase A(2) by Engineering Surface Charges

doi:10.1007/s12010-013-0437-2

CORC > 北京大学 > 化学与分子工程学院

	Tailoring the pH Dependence of Human Non-pancreatic Secretory Phospholipase A(2) by Engineering Surface Charges
	Qiu, Shunchen ; Lai, Luhua
刊名	应用生物化学与生物技术
	2013
关键词	Phospholipase A(2) Protein engineering Enzyme reaction optimum pH Catalytic mechanism The catalytic triad mechanism The calcium-coordinated oxyanion mechanism GROUP-IIA ACTIVE-SITE CATALYTIC MECHANISM SERINE-PROTEASE PANCREATIC PHOSPHOLIPASE-A2 CRYSTAL-STRUCTURE ENZYME CATALYSIS OPTIMUM BINDING STABILITY
DOI	10.1007/s12010-013-0437-2
英文摘要	Human non-pancreatic secretory phospholipase A(2) (hnpsPLA(2)) catalyzes the sn-2 acyl hydrolysis of phospholipids. It was reported that hnpsPLA(2) is involved in various diseases like inflammation, cancer, and so on. This enzyme also exhibits anti-bacterial and anti-virus activities. It is active over a broad pH range, with higher activity at alkaline conditions. In order to make it suitable as a potential bactericide, high activity at neutral pH is preferable. We have tried to tailor the pH dependence of hnpsPLA(2) activity by replacing its surface charged residues. Three surface charge replacements, Arg42Glu, Arg100Glu, and Glu89Lys, showed increased activities at neutral pH, which are 2.3, 2.8, and 2.3 times that of the wild-type enzyme at pH 7. Both the positive-to-negative and negative-to-positive mutations lowered the optimum enzymatic reaction pH of hnpsPLA(2), indicating that the enzyme pH profile depends on a delicate balance of charged residues. The activity changes are in good agreement with the recently proposed calcium-coordinated catalytic triad mechanism. This study also provides a general means of enhancing hnpsPLA(2) activity at low pH.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000326374700013&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; SCI(E); EI; PubMed; 1; ARTICLE; 6; 1454-1464; 171
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/149777]
专题	化学与分子工程学院生命科学学院
推荐引用方式 GB/T 7714	Qiu, Shunchen,Lai, Luhua. Tailoring the pH Dependence of Human Non-pancreatic Secretory Phospholipase A(2) by Engineering Surface Charges[J]. 应用生物化学与生物技术,2013.
APA	Qiu, Shunchen,&Lai, Luhua.(2013).Tailoring the pH Dependence of Human Non-pancreatic Secretory Phospholipase A(2) by Engineering Surface Charges.应用生物化学与生物技术.
MLA	Qiu, Shunchen,et al."Tailoring the pH Dependence of Human Non-pancreatic Secretory Phospholipase A(2) by Engineering Surface Charges".应用生物化学与生物技术 (2013).