Related Experiment Video
Updated: Jan 14, 2026
07:02
TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks
Published on: May 17, 2020
25.8K
Activity-based protein profiling of infected plants
Farnusch Kaschani1, Christian Gu, Renier A L van der Hoorn
1The Plant Chemetics Laboratory, Chemical Genomics Centre, Max Planck Institute for Plant Breeding Research, Cologne, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2011
Summary
Activity-based protein profiling (ABPP) uses chemical probes to label and identify active proteins in complex biological samples. This method allows for the comparison of protein activity across different proteomes, as demonstrated in infected Arabidopsis plants.
Area of Science:
- Biochemistry
- Proteomics
- Chemical Biology
Background:
- Activity-based protein profiling (ABPP) is a key technique for analyzing protein function.
- ABPP utilizes chemical probes that covalently bind to the active sites of enzymes.
- These probes enable the detection, purification, and identification of active proteins within a proteome.
Purpose of the Study:
- To provide a detailed protocol for ABPP, covering labeling, purification, and identification of proteins.
- To demonstrate the comparison of protein activity between two different proteomes.
- To illustrate the application of ABPP in identifying specific enzymes in plant-pathogen interactions.
Main Methods:
- Development and application of activity-based probes (e.g., trifunctional probe TriFP).
- Covalent labeling of active proteins via reaction with active site residues.
- Purification and separation of labeled proteins, often using gel electrophoresis.
- Mass spectrometry-based identification of labeled proteins.
Main Results:
- Successful labeling, purification, and identification of proteins using ABPP.
- Demonstration of differential protein activity between two proteomes.
- Identification of specific serine hydrolases in Arabidopsis plants infected with Botrytis cinerea.
Conclusions:
- ABPP is a robust method for comparing protein activity across proteomes.
- The described protocol facilitates the comprehensive analysis of enzyme activity.
- ABPP is valuable for studying plant-pathogen interactions and identifying key enzymes involved.
