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Updated: Jul 10, 2026
09:35
Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)
Published on: November 29, 2014
Comparative Real-Time Kinetics of Ligand-Receptor Interactions Using Immobilization-Based Sensing Readouts
Yazheng Wang1,2, Yalun Wu3,4, Lauren A Mayse1,5
1Department of Physics, Syracuse University, 201 Physics Building, Syracuse, New York, New York 13244, United States.
Analytical Chemistry
|July 9, 2026
Summary
Quantitative analysis of early ligand-EGFR binding kinetics reveals complex interactions. Immobilization techniques influence results, and glycosylation significantly impacts receptor affinity.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Signaling
Background:
- Receptor tyrosine kinase (RTK) and growth factor ligand interactions are vital for cellular communication.
- Early kinetic events in these signaling pathways are not fully quantified.
- Understanding ligand-receptor binding dynamics is crucial for deciphering cell signaling.
Purpose of the Study:
- To quantitatively analyze the real-time binding kinetics of high-affinity ligands with epidermal growth factor receptor (EGFR) isoforms.
- To investigate the influence of immobilization-based sensing techniques on kinetic and affinity measurements.
- To determine the role of glycosylation in ligand-EGFR interactions.
Main Methods:
- Employed biolayer interferometry (BLI) and surface plasmon resonance (SPR) for real-time binding kinetics analysis.
- Utilized high-affinity ligands and the full extracellular domain of various EGFR isoforms.
- Investigated interactions with both native and deglycosylated EGFR variants.
Main Results:
- Both BLI and SPR revealed complex binding kinetics with fast and slow dissociation phases, indicating two binding substates in ligand-EGFR interactions.
- Sensing technique choice significantly impacted kinetic parameters and relative ligand-EGFR interaction strengths.
- Glycan side chains at N151 did not affect interactions, but extensive deglycosylation markedly reduced ligand binding affinity.
Conclusions:
- Ligand-EGFR interactions exhibit complex kinetics, characterized by multiple binding substates.
- Immobilization strategies critically influence the quantitative assessment of these interactions.
- Post-translational modifications, specifically glycosylation, play a significant role in modulating ligand-receptor affinity.
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