Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells.
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2022-06
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Metabolites 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).
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Hsu, Che-Chia, Zhi-Gang Xu, Jie Lei, Zhong-Zhu Chen, Hong-Yu Li and Hui-Kuan Lin (2022). Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells. STAR protocols, 3(2). p. 101385. 10.1016/j.xpro.2022.101385 Retrieved from https://hdl.handle.net/10161/30659.
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Che-Chia Hsu
My research has focused on mitochondrial functions in cancer metabolism and understand the role of mitochondrial dynamics in cellular function and human diseases including cancers. Additionally, I also continuously dissect cancer metabolism and identifying potential metabolic vulnerabilities of cancer initiation, progression and metastasis using several in vitro, ex vivo and in vivo genetical approaches such as CRISPR/Cas9 knockout, mouse/ human organoid cultures and genetically engineered mouse models, thereby characterizing molecular mechanisms regulated by metabolic pathways and developing potential metabolic interventions for targeting cancers.
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