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Sortase-mediated chemical protein synthesis reveals the bidentate binding of bisphosphorylated p62 with K63 diubiquitin
Tan, Xiang-Long1,2; Pan, Man1; Zheng, Yong2,3; Gao, Shuai1; Liang, Lu-Jun1; Li, Yi-Ming2
2017-10-01
发表期刊CHEMICAL SCIENCE
卷号8期号:10页码:6881-6887
摘要Phosphorylation of S403 or S407 of the autophagic receptor protein p62 has recently been discovered to enhance the binding of p62 with ubiquitinated protein substrates to upregulate selective autophagy. To elucidate the molecular mechanism of how phosphorylation regulates the recruitment of ubiquitinated proteins, we report the first chemical synthesis of homogeneously phosphorylated p62, which enables the setting up of accurate in vitro systems for biochemical studies. Our synthesis employs the technology of sortase A-mediated protein hydrazide ligation, which successfully affords three types of phosphorylated p62 at the multi-milligram scale. Quantitative biochemical measurements show that the binding affinity of S403/S407-bisphosphorylated p62 to K63 diubiquitin is significantly higher than that of mono-phosphorylated p62. This finding suggests that phosphorylated S403 and S407 sites should bind to different epitopes on the ubiquitin chain. Furthermore, glutamate mutation is found to give a significantly impaired binding affinity, implying the necessity of using chemically synthesized phosphorylated p62 for the biochemical study of selective autophagy.
文章类型Article
WOS标题词Science & Technology ; Physical Sciences
资助者National Natural Science Foundation of China(21372058 ; National Natural Science Foundation of China(21372058 ; China Postdoctoral Science Foundation(2016M601999) ; China Postdoctoral Science Foundation(2016M601999) ; Fundamental Research Funds for the Central Universities ; Fundamental Research Funds for the Central Universities ; 21572043) ; 21572043) ; National Natural Science Foundation of China(21372058 ; National Natural Science Foundation of China(21372058 ; China Postdoctoral Science Foundation(2016M601999) ; China Postdoctoral Science Foundation(2016M601999) ; Fundamental Research Funds for the Central Universities ; Fundamental Research Funds for the Central Universities ; 21572043) ; 21572043)
DOI10.1039/c7sc02937c
关键词[WOS]SELECTIVE AUTOPHAGY ; PEPTIDE HYDRAZIDES ; CRYSTAL-STRUCTURE ; UBIQUITINATED PROTEINS ; LIGATION ; DEGRADATION ; RECOGNITION ; CHAINS ; DEUBIQUITINATION ; PHOSPHORYLATION
收录类别SCI
语种英语
资助者National Natural Science Foundation of China(21372058 ; National Natural Science Foundation of China(21372058 ; China Postdoctoral Science Foundation(2016M601999) ; China Postdoctoral Science Foundation(2016M601999) ; Fundamental Research Funds for the Central Universities ; Fundamental Research Funds for the Central Universities ; 21572043) ; 21572043) ; National Natural Science Foundation of China(21372058 ; National Natural Science Foundation of China(21372058 ; China Postdoctoral Science Foundation(2016M601999) ; China Postdoctoral Science Foundation(2016M601999) ; Fundamental Research Funds for the Central Universities ; Fundamental Research Funds for the Central Universities ; 21572043) ; 21572043)
WOS研究方向Chemistry
WOS类目Chemistry, Multidisciplinary
WOS记录号WOS:000411730500018
引用统计
被引频次:8[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.hfcas.ac.cn:8080/handle/334002/33740
专题中科院强磁场科学中心
作者单位1.Tsinghua Univ, Dept Chem, Key Lab Bioorgan Phosphorus Chem & Chem Biol, Tsinghua Peking Ctr Life Sci,Minist Educ, Beijing 100084, Peoples R China
2.Hefei Univ Technol, Sch Biol & Med Engn, Hefei 230009, Anhui, Peoples R China
3.Chinese Acad Sci, High Magnet Field Lab, Hefei 230031, Anhui, Peoples R China
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GB/T 7714
Tan, Xiang-Long,Pan, Man,Zheng, Yong,et al. Sortase-mediated chemical protein synthesis reveals the bidentate binding of bisphosphorylated p62 with K63 diubiquitin[J]. CHEMICAL SCIENCE,2017,8(10):6881-6887.
APA Tan, Xiang-Long,Pan, Man,Zheng, Yong,Gao, Shuai,Liang, Lu-Jun,&Li, Yi-Ming.(2017).Sortase-mediated chemical protein synthesis reveals the bidentate binding of bisphosphorylated p62 with K63 diubiquitin.CHEMICAL SCIENCE,8(10),6881-6887.
MLA Tan, Xiang-Long,et al."Sortase-mediated chemical protein synthesis reveals the bidentate binding of bisphosphorylated p62 with K63 diubiquitin".CHEMICAL SCIENCE 8.10(2017):6881-6887.
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