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

Publication ,  Journal Article
Hsu, C-C; Xu, Z-G; Lei, J; Chen, Z-Z; Li, H-Y; Lin, H-K
Published in: STAR Protoc
June 17, 2022

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).

Duke Scholars

Published In

STAR Protoc

DOI

EISSN

2666-1667

Publication Date

June 17, 2022

Volume

3

Issue

2

Start / End Page

101385

Location

United States

Related Subject Headings

  • Inositol
  • Cells, Cultured
  • Cell Line
  • Carrier Proteins
  • Biotin
 

Citation

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Hsu, C.-C., Xu, Z.-G., Lei, J., Chen, Z.-Z., Li, H.-Y., & Lin, H.-K. (2022). Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells. STAR Protoc, 3(2), 101385. https://doi.org/10.1016/j.xpro.2022.101385
Hsu, Che-Chia, Zhi-Gang Xu, Jie Lei, Zhong-Zhu Chen, Hong-Yu Li, and Hui-Kuan Lin. “Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells.STAR Protoc 3, no. 2 (June 17, 2022): 101385. https://doi.org/10.1016/j.xpro.2022.101385.
Hsu C-C, Xu Z-G, Lei J, Chen Z-Z, Li H-Y, Lin H-K. Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells. STAR Protoc. 2022 Jun 17;3(2):101385.
Hsu, Che-Chia, et al. “Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells.STAR Protoc, vol. 3, no. 2, June 2022, p. 101385. Pubmed, doi:10.1016/j.xpro.2022.101385.
Hsu C-C, Xu Z-G, Lei J, Chen Z-Z, Li H-Y, Lin H-K. Identification of myo-inositol-binding proteins by using the biotin pull-down strategy in cultured cells. STAR Protoc. 2022 Jun 17;3(2):101385.
Journal cover image

Published In

STAR Protoc

DOI

EISSN

2666-1667

Publication Date

June 17, 2022

Volume

3

Issue

2

Start / End Page

101385

Location

United States

Related Subject Headings

  • Inositol
  • Cells, Cultured
  • Cell Line
  • Carrier Proteins
  • Biotin