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Updated: Dec 11, 2025
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Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy NMR and Microscale Thermophoresis MST
Published on: November 2, 2018
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Detecting protein-protein interactions by Xe-129 NMR
Zhuangyu Zhao1, Benjamin W Roose1, Serge D Zemerov1
1Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. ivandmo@sas.upenn.edu.
Chemical Communications (Cambridge, England)
|August 21, 2020
Summary
This study introduces a novel protein complementation assay (PCA) using split TEM-1 β-lactamase for detecting protein-protein interactions (PPIs). The assay generates a sensitive 129Xe NMR signal upon protein interaction within bacterial cells.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophysics
Background:
- Current methods for detecting protein-protein interactions (PPIs) have limitations.
- The development of sensitive and versatile assays for PPI detection is crucial in biological research.
Purpose of the Study:
- To develop a novel bioanalytical method for detecting protein-protein interactions (PPIs).
- To utilize a split TEM-1 β-lactamase protein complementation assay (PCA) for sensitive detection of PPIs.
Main Methods:
- A split TEM-1 β-lactamase protein complementation assay (PCA) was employed.
- The interaction between cFos/cJun leucine zipper domains was used to reconstitute TEM-1.
- 129Xe NMR spectroscopy was utilized to detect the reconstituted enzyme activity in bacterial cells.
Main Results:
- Reconstitution of TEM-1 β-lactamase through protein interaction generated a sensitive 129Xe NMR signal.
- The assay successfully detected protein-protein interactions within a cellular environment.
Conclusions:
- The split TEM-1 β-lactamase PCA offers a sensitive method for detecting protein-protein interactions.
- This approach enables the detection of PPIs using 129Xe NMR in bacterial cells, overcoming limitations of existing methods.
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