Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage

doi:10.1038/cr.2014.4

KMS > 免疫化学研究所 > 特聘教授组 > Roger D. Kornberg组

	Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage
	Zhao, Lixia1,2 ; Hua, Tian 1; Crowley, Christopher 3; Ru, Heng 1; Ni, Xiangmin 1; Shaw, Neil 1; Jiao, Lianying 1; Ding, Wei 1; Qu, Lu 1; Hung, Li-Wei 4; Huang, Wei2 ; Liu, Lei 5; Ye, Keqiang 6; Ouyang, Songying 1; Cheng, Genhong 3; Liu, Zhi-Jie1,2
	2014-03
发表期刊	CELL RESEARCH (IF:28.1[JCR-2023],36.4[5-Year])
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	正在获取...
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.