Sortase-mediated chemical protein synthesis reveals the bidentate binding of bisphosphorylated p62 with K63 diubiquitin.
Sortase-mediated chemical protein synthesis reveals the bidentate binding of bisphosphorylated p62 with K63 diubiquitin.
复制标题
分选酶介导的化学蛋白质合成揭示了双磷酸化 p62 与 K63 双泛素的双齿结合
DOI：
10.1039/c7sc02937c
复制
发表时间：
2017-10-01
期刊：
影响因子：
8.4
通讯作者：
Li YM
中科院分区：
文献类型：
--
作者：
Tan XL;Pan M;Zheng Y;Gao S;Liang LJ;Li YM
This work reports the first chemical synthesis of the phosphorylated p62 protein and reveals a bidentate binding model of bisphosphorylated p62. 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.
登录
查看更多内容
影响因子：
15
作者：
Levinson AM;McGee JH;Roberts AG;Creech GS;Wang T;Peterson MT;Hendrickson RC;Verdine GL;Danishefsky SJ
通讯作者：
Danishefsky SJ
DOI：
10.1002/anie.201600638
发表时间：
2016-04-11
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Jbara M;Maity SK;Morgan M;Wolberger C;Brik A
通讯作者：
Brik A
DOI：
10.1002/cbic.201600276
发表时间：
2016-08-17
期刊：
Chembiochem : a European journal of chemical biology
影响因子：
--
作者：
Crowe SO;Pham GH;Ziegler JC;Deol KK;Guenette RG;Ge Y;Strieter ER
通讯作者：
Strieter ER
影响因子：
4.8
作者：
Hosing, Amol S.;Valerie, Nicholas C. K.;Larner, James M.
通讯作者：
Larner, James M.
影响因子：
56.9
作者：
DAWSON, PE;MUIR, TW;KENT, SBH
通讯作者：
KENT, SBH