Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage
Zhao, Lixia1,2; Hua, Tian1; Crowley, Christopher3; Ru, Heng1; Ni, Xiangmin1; Shaw, Neil1; Jiao, Lianying1; Ding, Wei1; Qu, Lu1; Hung, Li-Wei4; Huang, Wei2; Liu, Lei5; Ye, Keqiang6; Ouyang, Songying1; Cheng, Genhong3; Liu, Zhi-Jie1,2
2014-03
Source PublicationCELL RESEARCH
ISSN1001-0602
Volume24Issue:3Pages:344-358
Status已发表
DOI10.1038/cr.2014.4
AbstractAsparaginyl 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.
Keywordasparaginyl endopeptidase autoproteolytic maturation crystal structure innate immunity
Indexed BySCI ; CSCD
Language英语
Funding ProjectNational 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 Research AreaCell Biology
WOS SubjectCell Biology
WOS IDWOS:000332246500010
CSCD IDCSCD:5093932
PublisherINST BIOCHEMISTRY & CELL BIOLOGY
WOS KeywordMAMMALIAN LEGUMAIN ; MACROMOLECULAR STRUCTURES ; CYSTEINE PROTEASES ; ANTIGEN ; EXPRESSION ; CLEAVAGE ; PROTEIN ; DIFFRACTION ; INHIBITION ; CYSTATINS
Original Document TypeArticle
Citation statistics
Cited Times:61[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttps://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1071
Collection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
生命科学与技术学院_硕士生
物质科学与技术学院_硕士生
iHuman研究所_PI研究组_华甜组
Corresponding AuthorLiu, Zhi-Jie
Affiliation1.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
First Author AffilicationiHuman Institute
Corresponding Author AffilicationiHuman Institute
Recommended Citation
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.
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