Single-Molecule Enzymatic Reaction Dynamics
EMSL Project ID
3277
Abstract
T4 lysozyme catalyzes the hydrolization of the bacterial cell wall and involves complex conformational changes. Although this hydrolization is one of the most important enzymatic reactions, its mechanism is still largely unknown. Conventional enzymatic reaction assay has not able to resolve the complex reaction mechanism. Enzymatic reactions often involve complex environment and mechanism, therefore, it is often crucial to probe directly the specific conformational changes involved in an enzymatic reaction. The hinge-bending motions associated with the T4 lysozyme hydrolysis enzymatic reaction are critical but cannot be studied by either conventional ensemble-averaged measurements or the above mentioned single-molecule spectroscopic approaches. In this project, we have used intramolecular fluorescence resonance energy transfer between a single pair of dye labeled on T4 lysozyme to probe the conformational change during enzymatic reaction. Our approach has the advantage of characterizing the enzymatic-activity-related conformational fluctuation, although the product release is not directly probed.
Project Details
Project type
Exploratory Research
Start Date
2003-05-13
End Date
2004-05-20
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members