Template-assisted rational design of peptide inhibitors of furin using the lysine fragment of the mung bean trypsin inhibitor
文献类型：期刊论文
| 作者 | Tao, H; Zhang, Z; Shi, J; Shao, XX; Cui, D; Chi, CW |
| 刊名 | FEBS JOURNAL
 |
| 出版日期 | 2006 |
| 卷号 | 273期号:17页码:3907-3914 |
| 关键词 | furin kexin molecular design mung bean trypsin inhibitor peptide synthesis |
| 通讯作者 | Chi, CW (reprint author), Chinese Acad Sci, Grad Sch Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Inst Biochem & Cell Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,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. |
| 学科主题 | Biochemistry & Molecular Biology |
| 类目[WOS] | Biochemistry & Molecular Biology |
| 关键词[WOS] | CRYSTAL-STRUCTURE ; KEX2 PROTEASE ; PROPROTEIN CONVERTASES ; PROTEINASE-INHIBITOR ; ANGSTROM RESOLUTION ; SUNFLOWER SEEDS ; SPECIFICITY ; ACTIVATION ; ENDOPROTEASE ; PRECURSOR |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000239858300004 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1790]  |
| 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 | Tao, H,Zhang, Z,Shi, J,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, H,Zhang, Z,Shi, J,Shao, XX,Cui, D,&Chi, CW.(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, H,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. |
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