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Published on: February 5, 2022
A proteome-wide map of 20(S)-hydroxycholesterol interactors in cell membranes
Yu-Shiuan Cheng1, Tianyi Zhang1, Xiang Ma1
1Department of Chemistry, Division of Chemistry & Chemical Engineering, California Institute of Technology, Pasadena, CA, USA.
Researchers developed a new chemoproteomics probe to map cellular interactions of 20(S)-hydroxycholesterol (20(S)-OHC). This study identifies key protein targets involved in immune response and cancer, revealing new therapeutic strategies.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Oxysterols (OHCs) are hydroxylated cholesterol metabolites with critical roles in cellular functions and disease pathogenesis.
- Analyzing live-cell, proteome-wide OHC interactions is challenging due to their non-covalent binding and membrane partitioning.
Purpose of the Study:
- To develop a chemoproteomics probe for mapping the proteome-wide interactions of 20(S)-hydroxycholesterol (20(S)-OHC) in living cells.
- To identify OHC-interacting proteins and elucidate their roles in biological processes.
Main Methods:
- Development of a structurally precise chemoproteomics probe for 20(S)-OHC.
- Proteome-wide target identification in live-cell membranes.
- Competition experiments to determine ligand selectivity and binding site reconstruction.
Main Results:
- A comprehensive map of 20(S)-OHC proteome-wide targets in cellular membranes was generated.
- OHC-interacting proteins were found to be enriched in pathways related to immune response and cancer.
- 20(S)-OHC was identified as a chemo-, regio-, and stereoselective ligand for transmembrane protein 97 (Tmem97/the σ2 receptor).
- The binding site of 20(S)-OHC within Tmem97 was reconstructed.
Conclusions:
- Multiplexed, quantitative analysis of cellular target engagement reveals novel aspects of metabolite activity.
- The study identifies Tmem97 as a key OHC-binding protein, offering potential for molecular therapy development.
- This approach provides a framework for exploring metabolite-protein interactions and discovering new therapeutic targets.
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