Browsing by Author "Xu, Zhi-Gang"
Now showing 1 - 3 of 3
- Results Per Page
- Sort Options
Item Open Access Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells.(STAR protocols, 2022-06) Hsu, Che-Chia; Xu, Zhi-Gang; Lei, Jie; Chen, Zhong-Zhu; Li, Hong-Yu; Lin, Hui-KuanMetabolites are not only substrates in metabolic reactions, but they also serve as signaling molecules to regulate diverse biological functions. Identification of the binding proteins for the metabolites helps in the understanding of their functions beyond the classic metabolic pathways in which they are involved. We provide the protocol for synthesizing the biotin-labeled myo-inositol, which is used to identify its binding proteins by using biotin pull-down assay, given there is no available tool for the rapid screening of inositol-binding proteins in cells and in vitro systems. Biotin-labeled inositol probe therefore provides a tool to identify inositol's sensors. For complete details on the use and execution of this protocol, please refer to Hsu et al. (2021).Item Open Access Inositol serves as a natural inhibitor of mitochondrial fission by directly targeting AMPK.(Molecular cell, 2021-09) Hsu, Che-Chia; Zhang, Xian; Wang, Guihua; Zhang, Weina; Cai, Zhen; Pan, Bo-Syong; Gu, Haiwei; Xu, Chuan; Jin, Guoxiang; Xu, Xiangshang; Manne, Rajesh Kumar; Jin, Yan; Yan, Wei; Shao, Jingwei; Chen, Tingjin; Lin, Emily; Ketkar, Amit; Eoff, Robert; Xu, Zhi-Gang; Chen, Zhong-Zhu; Li, Hong-Yu; Lin, Hui-KuanMitochondrial dynamics regulated by mitochondrial fusion and fission maintain mitochondrial functions, whose alterations underline various human diseases. Here, we show that inositol is a critical metabolite directly restricting AMPK-dependent mitochondrial fission independently of its classical mode as a precursor for phosphoinositide generation. Inositol decline by IMPA1/2 deficiency elicits AMPK activation and mitochondrial fission without affecting ATP level, whereas inositol accumulation prevents AMPK-dependent mitochondrial fission. Metabolic stress or mitochondrial damage causes inositol decline in cells and mice to elicit AMPK-dependent mitochondrial fission. Inositol directly binds to AMPKγ and competes with AMP for AMPKγ binding, leading to restriction of AMPK activation and mitochondrial fission. Our study suggests that the AMP/inositol ratio is a critical determinant for AMPK activation and establishes a model in which AMPK activation requires inositol decline to release AMPKγ for AMP binding. Hence, AMPK is an inositol sensor, whose inactivation by inositol serves as a mechanism to restrict mitochondrial fission.Item Open Access Lactate Is a Natural Suppressor of RLR Signaling by Targeting MAVS(Cell, 2019-06) Zhang, Weina; Wang, Guihua; Xu, Zhi-Gang; Tu, Haiqing; Hu, Fuqing; Dai, Jiang; Chang, Yan; Chen, Yaqi; Lu, Yanjun; Zeng, Haolong; Cai, Zhen; Han, Fei; Xu, Chuan; Jin, Guoxiang; Sun, Li; Pan, Bo-Syong; Lai, Shiue-Wei; Hsu, Che-Chia; Xu, Jia; Chen, Zhong-Zhu; Li, Hong-Yu; Seth, Pankaj; Hu, Junbo; Zhang, Xuemin; Li, Huiyan; Lin, Hui-Kuan