Template-assisted rational design of peptide inhibitors of furin using the lysine fragment of the mung bean trypsin inhibitor

doi:10.1111/j.1742-4658.2006.05393.x

CORC > 金属研究所 > 中国科学院金属研究所

	Template-assisted rational design of peptide inhibitors of furin using the lysine fragment of the mung bean trypsin inhibitor
	Tao, Hu ; Zhang, Zhen ; Shi, Jiahao ; Shao, Xiao-xia ; Cui, Dafu ; Chi, Cheng-Wu
刊名	FEBS JOURNAL
	2006-09-01
卷号	273 期号:17 页码:3907-3914
关键词	furin kexin molecular design mung bean trypsin inhibitor peptide synthesis
ISSN号	1742-464X
DOI	10.1111/j.1742-4658.2006.05393.x
通讯作者	Chi, Cheng-Wu(chi@sunm.shcnc.ac.cn)
英文摘要	Highly active, small-molecule furin inhibitors are attractive drug candidates to fend off bacterial exotoxins and viral infection. Based on the 22-residue, active Lys fragment of the mung bean trypsin inhibitor, a series of furin inhibitors were designed and synthesized, and their inhibitory activity towards furin and kexin was evaluated using enzyme kinetic analysis. The most potent inhibitor, containing 16 amino acid residues with a K-i value of 2.45 x 10(-9) M for furin and of 5.60 x 10(-7) M for kexin, was designed with three incremental approaches. First, two nonessential Cys residues in the Lys fragment were deleted via a Cys-to-Ser mutation to minimize peptide misfolding. Second, residues in the reactive site of the inhibitor were replaced by the consensus substrate recognition sequence of furin, namely, Arg at P-1, Lys at P-2, Arg at P-4 and Arg at P-6. In addition, the P-7 residue Asp was substituted with Ala to avoid possible electrostatic interference with furin inhibition. Finally, the extra N-terminal and C-terminal residues beyond the doubly conjugated disulfide loops were further truncated. However, all resultant synthetic peptides were found to be temporary inhibitors of furin and kexin during a prolonged incubation, with the scissile peptide bond between P-1 and P-1' being cleaved to different extents by the enzymes. To enhance proteolytic resistance, the P-1' residue Ser was mutated to D-Ser or N-methyl-Ser. The N-methyl-Ser mutant gave rise to a K-i value of 4.70 x 10(-8) M for furin, and retained over 80% inhibitory activity even after a 3 h incubation with the enzyme. By contrast, the D-Ser mutant was resistant to cleavage, although its inhibitory activity against furin drastically decreased. Our findings identify a useful template for the design of potent, specific and stable peptide inhibitors of furin, shedding light on the molecular determinants that dictate the inhibition of furin and kexin.
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	BLACKWELL PUBLISHING
WOS记录号	WOS:000239858300004
内容类型	期刊论文
源URL	[http://ir.imr.ac.cn/handle/321006/85853]
专题	金属研究所_中国科学院金属研究所
通讯作者	Chi, Cheng-Wu
作者单位	1.Tongji Univ, Inst Prot Res, Shanghai, Peoples R China 2.Chinese Acad Sci, Grad Sch Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China
推荐引用方式 GB/T 7714	Tao, Hu,Zhang, Zhen,Shi, Jiahao,et al. Template-assisted rational design of peptide inhibitors of furin using the lysine fragment of the mung bean trypsin inhibitor[J]. FEBS JOURNAL,2006,273(17):3907-3914.
APA	Tao, Hu,Zhang, Zhen,Shi, Jiahao,Shao, Xiao-xia,Cui, Dafu,&Chi, Cheng-Wu.(2006).Template-assisted rational design of peptide inhibitors of furin using the lysine fragment of the mung bean trypsin inhibitor.FEBS JOURNAL,273(17),3907-3914.
MLA	Tao, Hu,et al."Template-assisted rational design of peptide inhibitors of furin using the lysine fragment of the mung bean trypsin inhibitor".FEBS JOURNAL 273.17(2006):3907-3914.