The inhibitory activities of recombinant eglin C mutants on kexin and furin, using site-directed mutagenesis and molecular modeling

	The inhibitory activities of recombinant eglin C mutants on kexin and furin, using site-directed mutagenesis and molecular modeling
	Fei, H; Luo, MJ; Ye, YZ; Ding, DF; Chi, CW
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2001
卷号	33 期号:6 页码:591-599
关键词	proprotein convertase proteinase inhibitor site-directed mutagenesis molecular modeling
通讯作者	,
英文摘要	Mammalian furin and yeast kexin are members of the proprotein convertase family involved in the proteolytic processing of many important precursor proteins. Here the gene coding for the subtilisin inhibitor eglin C was totally synthesized and expressed in E. coli. Substitution of residues at each position P-1, P-2 and P-4 of eglin C with a basic residue using protein engineering could make eglin C a very strong inhibitor for furin (K-i around 10(-9) mol/L), and even more strong for kexin (K-i around 10(-11) mol/L). Results indicated that: (1) A basic residue Lys or Arg at P-1 site is prerequisite for the inhibitor. (2) The second mutation with basic residue at P-4 site drastically increase the inhibitory activity by two orders of magnitude. (3) A basic residue at P-2 site is favorable for the binding to the enzyme, but unfavorable for the stability of the inhibitor, resulting in a temporary inhibition. (4) A hydrophobic residue is preferential at P-3 site. Based on the known crystal structures of subtilisin and eglin C, the interaction between the enzyme and inhibitor was modeled, and their involved residues were predicted which gave a good explanation to the experimental results.
学科主题	Biochemistry & Molecular Biology ; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	PROPROTEIN CONVERTASES ; SPECIFICITY ; ENDOPROTEASE ; ACTIVATION ; CLEAVAGE ; PROTEASE ; GENE ; GLYCOPROTEIN ; PRECURSOR ; PEPTIDES
收录类别	SCI
语种	英语
WOS记录号	WOS:000172362600001
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2683]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Fei, H,Luo, MJ,Ye, YZ,et al. The inhibitory activities of recombinant eglin C mutants on kexin and furin, using site-directed mutagenesis and molecular modeling[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2001,33(6):591-599.
APA	Fei, H,Luo, MJ,Ye, YZ,Ding, DF,&Chi, CW.(2001).The inhibitory activities of recombinant eglin C mutants on kexin and furin, using site-directed mutagenesis and molecular modeling.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,33(6),591-599.
MLA	Fei, H,et al."The inhibitory activities of recombinant eglin C mutants on kexin and furin, using site-directed mutagenesis and molecular modeling".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 33.6(2001):591-599.