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Single molecule recordings of lysozyme activity
Yongki Choi1, Gregory A Weiss, Philip G Collins
1Department of Physics and Astronomy, University of California at Irvine, Irvine, California 92697, USA.
Physical Chemistry Chemical Physics : PCCP
|June 12, 2013
Summary
Single molecule bioelectronic circuits reveal T4 lysozyme
Area of Science:
- Biophysics
- Biochemistry
- Enzyme kinetics
Background:
- Single-molecule techniques offer high-resolution insights into biological processes.
- Understanding enzyme mechanisms at the molecular level is crucial for drug development and biotechnology.
Purpose of the Study:
- To utilize single molecule bioelectronic circuits to investigate the catalytic mechanism and kinetic variability of T4 lysozyme.
- To demonstrate the capability of bioelectronic circuits for studying enzyme activity with bond-by-bond resolution.
Main Methods:
- Fabrication and utilization of single molecule bioelectronic circuits.
- Monitoring changes in electrical conductance to track individual T4 lysozyme molecules.
- Synthesizing and testing ten T4 lysozyme variants.
Main Results:
- T4 lysozyme was identified as a processive enzyme governed by 9 independent time constants.
- Variations in time constants with pH and substrate crosslinking provided insights into catalytic activity and dynamic disorder.
- A single amino acid was sufficient for effective signal generation in the circuits.
Conclusions:
- Single molecule bioelectronic circuits can elucidate previously invisible aspects of enzyme catalysis.
- The technique is sensitive to environmental factors like pH and substrate modifications.
- The bioelectronic circuit approach is versatile and extendable to other proteins for studying chemical kinetics.
