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Updated: Jun 27, 2026
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Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
Published on: October 15, 2016
Linking phospholipase mobility to activity by single-molecule wide-field microscopy
Susana Rocha1, James A Hutchison, Kalina Peneva
1Division of Molecular and Nano Materials, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001 Leuven, Belgium.
Summary
Single-molecule microscopy reveals how phospholipase enzyme mobility relates to their catalytic activity on cell membranes. This technique offers new insights into interfacial enzymatic catalysis and diffusion processes.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Cell membrane enzymatic reactions are vital for cellular functions.
- Interfacial enzymatic catalysis mechanisms, including diffusion, are not fully understood.
- Studying enzyme-substrate interactions at interfaces is challenging.
Purpose of the Study:
- To investigate interfacial enzymatic catalysis using single-molecule fluorescence microscopy.
- To correlate enzyme mobility with catalytic activity.
- To visualize substrate structural changes during enzymatic reactions.
Main Methods:
- Single-molecule wide-field fluorescence microscopy was employed.
- Phospholipase enzymes acting on phospholipid bilayers were imaged.
- Diffusion of individual enzymes and substrate structural changes were tracked simultaneously.
Main Results:
- The mobility of different phospholipase enzyme types was successfully correlated with their catalytic activity.
- Thousands of individual enzyme trajectories provided detailed mechanistic insights.
- Simultaneous visualization of enzyme diffusion and substrate modification was achieved.
Conclusions:
- Single-molecule microscopy provides novel information on interfacial enzymatic catalysis.
- The developed method is a promising approach for studying heterogeneous catalysis.
- Understanding enzyme mobility is key to understanding catalytic efficiency at interfaces.
