Split-TurboID-Based Proximity Labeling for Probing the Composition of Spatiotemporally Defined Protein Complexes In Planta.

Zhiyan Wen, Xinyu Zhang, Yuexiao Li, Dingliang Zhang, Savithramma P Dinesh-Kumar, Yongliang Zhang

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Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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6 authors.

Zhiyan WenState Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing, China.

Xinyu ZhangState Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing, China.

Yuexiao LiState Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing, China.

Dingliang ZhangState Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing, China.

Savithramma P Dinesh-KumarDepartment of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, USA. spdineshkumar@ucdavis.edu.

Yongliang ZhangState Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing, China. cauzhangyl@cau.edu.cn.

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6 · The paper itself

Abstract

Proteins frequently interact with other proteins to execute specific biological processes. Identifying protein-protein interactions (PPIs) is essential for understanding the molecular pathways underlying various physiological functions. Proximity labeling (PL), particularly the TurboID-based PL approach, has been widely utilized to identify PPIs across different organisms in recent years. For the first time, we applied the TurboID-based PL method to detect PPIs in plants. We recently demonstrated that the TurboID complementation approach, referred to as split-TurboID, can be used to screen for binding partners of defined protein complexes in planta. Here, we provide a detailed protocol for performing split-TurboID-based PL assays in Nicotiana benthamiana plants.

Indexed as

Multiprotein ComplexesNicotianaPlant ProteinsProtein Interaction MappingStaining and LabelingChromatography, LiquidHSP90 Heat-Shock ProteinsProtein BindingTandem Mass SpectrometryHSP90 Heat-Shock ProteinsMultiprotein ComplexesPlant ProteinsLC-MS/MSProtein-protein interactions (PPIs)Proximity labelingSGT1-HSP90 protein complexSplit-TurboID

Identifiers

PMID41642522

DOI10.1007/978-1-59745-198-7_4

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