An associate professor of biophysics from Clemson University, Hugo Sanabria, teamed with researchers from Heinrich Heine University in Germany to develop new optical imaging techniques that can monitor a single molecule.
Sanabria and his colleagues used Forster Resonance Energy Transfer (FRET) (also called Fluorescence Resonance Energy Transfer) and other tools to monitor the T4 Lysozyme (T4L) in action. The team’s findings, “Resolving dynamics and function of transient states in single enzyme molecules” are published in the March 6, 2020 edition of Nature Communications.
This FRET technique can help scientists gain a better understanding of how molecules function, and that’s important to developing structure-guided drugs.
The researchers used a FRET-based microscope to visualize biomolecules as small as a few nanometers. They put two fluorescent markers on a set of molecules and used the location of these markers to collect a set of distances. Using different locations on each molecule helped the researchers to be able to determine the shape of the molecule and how it moves.
In the study, Sanabria and his team examined lysozyme, which is an enzyme that breaks down the cell walls of certain bacteria. Lysozyme can be found in tears, mucus, and egg whites. What Sanabria found is that the structure of lysozyme was different than they thought. Sanabria says, “For the longest time, this molecule was considered a two-state molecule because of how it receives the substrate or cell wall of the target bacteria. However, we have identified a new functional state.”
Sanabria’s team is creating a database of FRET-based structural models that can be accessed by other researchers. Sanabria also wants to examine other molecules using FRET technology. According to Sanabria, “This optical method can be used to study protein folding and misfolding or any structural organization of biomolecules. It can also be used for drug screening and development, which requires knowing what a biomolecule looks like in order for a drug to target it.”