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Updated: Jun 13, 2026
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TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks
Published on: May 17, 2020
Screening and identification of protein-protein interaction using proximity labeling
Jiang Hua1, Francis Ka-Ming Chan2
1Liangzhu Laboratory, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, P.R. China.
Methods in Cell Biology
|June 11, 2026
Summary
This study introduces a proximity labeling protocol to identify new regulators of programmed cell death. The method maps interactors of key necroptosis proteins, advancing our understanding of cell death mechanisms.
Area of Science:
- Cellular Biology
- Molecular Biology
- Biochemistry
Background:
- Programmed cell death, including apoptosis, necroptosis, and pyroptosis, is vital in physiology and pathology.
- Necroptosis signaling involves protein-protein interactions, with mediators like RIPK1, RIPK3, and MLKL regulated by post-translational modifications.
- Identifying transient protein associations is crucial for understanding necroptosis regulation.
Purpose of the Study:
- To develop and present a detailed protocol for proximity labeling to map protein interactors.
- To identify novel regulators and transient interactors of key necroptosis mediators.
- To enhance the understanding of necroptosis signaling pathways.
Main Methods:
- Utilizing TurboID-based proximity labeling technology.
- Coupling proximity labeling with mass spectrometry for protein identification.
- Applying the protocol to map interactors of RIPK1, RIPK3, MLKL, and ZBP1.
Main Results:
- The protocol enables the identification of proteins in close proximity (∼10-20 nm) to target proteins.
- This method is effective for discovering weak and transient protein-protein interactions.
- The chapter provides a step-by-step guide for implementing this technique.
Conclusions:
- Proximity labeling is a powerful tool for dissecting complex signaling pathways like necroptosis.
- This protocol facilitates the discovery of new components involved in programmed cell death.
- Further research can leverage this method to explore other cell death modalities and signaling networks.
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