Cholesterylation of Smoothened is a calcium-accelerated autoreaction involving an intramolecular ester intermediate  

作　　者：Ao Hu Jing-Zan Zhang Jie Wang Chen-Chen Li Meng Yuan Gang Deng Zi-Cun Lin Zhi-Ping Qiu Hu-Yue Liu Xian-Wei Wang Peng-Cheng Wei Xiao He Xiaolu Zhao Wen-Wei Qiu Bao-Liang Song 

机构地区：[1]The Institute for Advanced Studies,Hubei Key Laboratory of Cell Homeostasis,College of Life Sciences,TaiKang Center for Life and Medical Sciences,TaiKang Medical School,Wuhan University,Wuhan,Hubei,China [2]Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development,School of Chemistry and Molecular Engineering,East China Normal University,Shanghai,China [3]College of Science,Zhejiang University of Technology,Hangzhou,Zhejiang,China [4]NYU-ECNU Center for Computational Chemistry at NYU Shanghai,Shanghai,China

出　　处：《Cell Research》2022年第3期288-301,共14页细胞研究（英文版）

基　　金：We thank Ms.Dan Liang for technical assitance,Dr.Wei Qi(ShanghaiTech University)for revising the manuscript.We appreciate the helpful discussion with Drs.Guo-Yin Yin Wuhan University)and Yun Zhao(Shanghail Istitutes for Biological Sciences);This work was supported by grants from the National Natural Science Foundation of China(22077035,91753204,31690102,91957103);Ministry of Science and Technology of China(2018YF A0800700.2019YFA0802701);Fountain-Valley Life Sciences Fund of University of Chinese Academy of Sciences Education Foundation.B.-L.S.acknowledges the support from the Tencent Foundation through the XPLORER PRIZE.

摘　　要：Hedgehog(Hh)is a morphogen that binds to its receptor Patched 1 and activates Smoothened(SMO),thereby governing embryonic development and postnatal tissue homeostasis.Cholesterol can bind and covalently conjugate to the luminal cysteine-rich domain(CRD)of human SMO at the D95 residue(D99 in mouse).The reaction mechanism and biological function of SMO cholesterylation have not been elucidated.Here,we show that the SMO-CRD undergoes auto cholesterylation which is boosted by calcium and involves an intramolecular ester intermediate.In cells,Hh stimulation elevates local calcium concentration in the SMO-localized endosomes through store-operated calcium entry.In addition,we identify the signaling-incompetent SMO D95E mutation,and the D95E mutant SMO can bind cholesterol but cannot be modifed or activated by cholesterol.The homozygous Smo^(O99E/D99E) knockin mice are embryonic lethal with severe developmental delay,demonstrating that cholesterylation of CRD is required for full-length SMO activation.Our work reveals the unique autocatalytic mechanism of SMO cholesterylation and an unprecedented role of calcium in Hh signaling.

关 键 词：SMOOTHENED CHOLESTEROL STIMULATION 

分 类 号：Q2[生物学—细胞生物学]