Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells

Che Chia Hsu, Zhi Gang Xu, Jie Lei, Zhong Zhu Chen, Hong Yu Li, Hui Kuan Lin

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

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).

Original languageEnglish (US)
Article number101385
JournalSTAR Protocols
Volume3
Issue number2
DOIs
StatePublished - Jun 17 2022
Externally publishedYes

Keywords

  • Biotechnology and
  • Cell culture
  • Cell-based Assays
  • Chemistry
  • Metabolism
  • Molecular Biology
  • Molecular/Chemical Probes
  • Protein Biochemistry
  • bioengineering

ASJC Scopus subject areas

  • General Neuroscience
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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