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TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks
Published on: May 17, 2020
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TurboID-Based Proximity Labeling: A Method to Decipher Protein-Protein Interactions in Plants
Yuanyuan Li1, Yongliang Zhang2, Savithramma P Dinesh-Kumar3
1Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|November 21, 2023
Summary
TurboID proximity labeling (PL) offers a faster, room-temperature method for studying plant protein-protein interactions (PPIs). This optimized protocol overcomes limitations of older techniques, enabling efficient analysis of plant cellular processes.
Area of Science:
- Plant molecular biology
- Biochemistry
- Proteomics
Background:
- Protein-protein interactions (PPIs) are crucial for cellular processes.
- Traditional PPI analysis methods have limitations in plant systems, including toxicity and long incubation times.
- Enzyme-catalyzed proximity labeling (PL) techniques offer alternatives but require optimization for plants.
Purpose of the Study:
- To describe an optimized protocol for TurboID-based proximity labeling (PL) in plants for studying PPIs.
- To demonstrate the efficacy of TurboID-PL compared to BioID in labeling endogenous plant proteins.
- To provide a adaptable method for investigating PPI networks in various plant species.
Main Methods:
- Agrobacterium-based transient expression of proteins.
- Biotin treatment and subsequent protein extraction.
- Removal of free biotin, quantification, and affinity purification of biotinylated proteins.
- Utilized TurboID-PL with a plant viral immune receptor (NLR class) as a model system.
Main Results:
- TurboID-based PL demonstrated superior performance over BioID in labeling endogenous proteins in plants.
- The optimized protocol enables rapid proximity labeling at room temperature.
- Successful application of the method for studying PPIs involving a plant immune receptor.
Conclusions:
- TurboID-based PL is an efficient and versatile tool for studying plant PPIs.
- The described protocol overcomes previous limitations, facilitating research in plant molecular biology.
- This method can be readily adapted for diverse plant species and PPI network analyses.
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