ShanghaiTech University Knowledge Management System
| Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage | |
| 2014-03 | |
| 发表期刊 | CELL RESEARCH
 |
| ISSN | 1001-0602 |
| 卷号 | 24期号:3页码:344-358 |
| 发表状态 | 已发表 |
| DOI | 10.1038/cr.2014.4 |
| 摘要 | Asparaginyl endopeptidase (AEP) is an endo/lysosomal cysteine endopeptidase with a preference for an asparagine residue at the P1 site and plays an important role in the maturation of toll-like receptors 3/7/9. AEP is known to undergo autoproteolytic maturation at acidic pH for catalytic activation. Here, we describe crystal structures of the AEP proenzyme and the mature forms of AEP. Structural comparisons between AEP and caspases revealed similarities in the composition of key residues and in the catalytic mechanism. Mutagenesis studies identified N44, R46, H150, E189, C191, S217/S218 and D233 as residues that are essential for the cleavage of the peptide substrate. During maturation, autoproteolytic cleavage of AEP's cap domain opens up access to the active site on the core domain. Unexpectedly, an intermediate autoproteolytic maturation stage was discovered at approximately pH 4.5 in which the partially activated AEP could be reversed back to its proenzyme form. This unique feature was confirmed by the crystal structure of AEPpH4.5 (AEP was matured at pH 4.5 and crystallized at pH 8.5), in which the broken peptide bonds were religated and the structure was transformed back to its proenzyme form. Additionally, the AEP inhibitor cystatin C could be digested by the fully activated AEP, but could not be digested by activated cathepsins. Thus, we demonstrate for the first time that cystatins may regulate the activity of AEP through substrate competition for the active site. |
| 关键词 | asparaginyl endopeptidase autoproteolytic maturation crystal structure innate immunity |
| 收录类别 | SCI ; CSCD |
| 语种 | 英语 |
| 资助项目 | National Natural Science Foundation of China[31330019] ; National Natural Science Foundation of China[31200559] ; National Natural Science Foundation of China[91313301] ; National Natural Science Foundation of China[31300613] |
| WOS研究方向 | Cell Biology |
| WOS类目 | Cell Biology |
| WOS记录号 | WOS:000332246500010 |
| CSCD记录号 | CSCD:5093932 |
| 出版者 | INST BIOCHEMISTRY & CELL BIOLOGY |
| WOS关键词 | MAMMALIAN LEGUMAIN ; MACROMOLECULAR STRUCTURES ; CYSTEINE PROTEASES ; ANTIGEN ; EXPRESSION ; CLEAVAGE ; PROTEIN ; DIFFRACTION ; INHIBITION ; CYSTATINS |
| 原始文献类型 | Article |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1071 |
| 专题 | 免疫化学研究所_特聘教授组_结构生物化学实验室 iHuman研究所 iHuman研究所_PI研究组_刘志杰组 iHuman研究所_PI研究组_程建军组 |
| 通讯作者 | Liu, Zhi-Jie |
| 作者单位 | 1.Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China 2.ShanghaiTech Univ, iHuman Inst, Shanghai 201210, Peoples R China 3.Univ Calif Los Angeles, Dept Microbiol Immunol & Mol Genet, Los Angeles, CA 90095 USA 4.Los Alamos Natl Lab, Div Phys, Los Alamos, NM 87545 USA 5.Tsinghua Univ, Dept Chem, Key Lab Bioorgan Phosphorus Chem & Chem Biol, Minist Educ, Beijing 100084, Peoples R China 6.Emory Univ, Sch Med, Dept Pathol & Lab Med, Atlanta, GA 30322 USA |
| 第一作者单位 | iHuman研究所 |
| 通讯作者单位 | iHuman研究所 |
| 推荐引用方式 GB/T 7714 | Zhao, Lixia,Hua, Tian,Crowley, Christopher,et al. Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage[J]. CELL RESEARCH,2014,24(3):344-358. |
| APA | Zhao, Lixia.,Hua, Tian.,Crowley, Christopher.,Ru, Heng.,Ni, Xiangmin.,...&Liu, Zhi-Jie.(2014).Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage.CELL RESEARCH,24(3),344-358. |
| MLA | Zhao, Lixia,et al."Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage".CELL RESEARCH 24.3(2014):344-358. |
| 条目包含的文件 | 下载所有文件 | |||||
| 文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
修改评论
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。