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

IMR OpenIR

	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
通讯作者	Chi, Cheng-Wu(chi@sunm.shcnc.ac.cn)
	2006-09-01
发表期刊	FEBS JOURNAL
ISSN	1742-464X
卷号	273 期号:17 页码:3907-3914
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.
关键词	furin kexin molecular design mung bean trypsin inhibitor peptide synthesis
DOI	10.1111/j.1742-4658.2006.05393.x
收录类别	SCI
语种	英语
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000239858300004
出版者	BLACKWELL PUBLISHING
引用统计	被引频次：1[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.imr.ac.cn/handle/321006/85854
专题	中国科学院金属研究所
通讯作者	Chi, Cheng-Wu
作者单位	1.Chinese Acad Sci, Grad Sch Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China 2.Tongji Univ, Inst Prot Res, Shanghai, 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.