Enzyme functional dynamics

Enzyme Functional Dynamics. In collaboration with Emil Pai (UofT, Biochemistry) we are investigating the dynamics of a fascinating bacterial enzyme, consisting of two 300 residue monomeric domains, and whose job is to rip apart a CF bond in a fluoroacetate species. We are able to observe the entire dynamics associated with a key indole species in the protein which coordinates with the substrate, while a second residue is involved in sweeping out the product from the active site. By labeling the protein with fluorotryptophan we can simultaneously monitor dynamics of all 9 Trp residues in the susbtrate-free state, and substrate or inhibitor loaded state. The data thus far provides insight into allostery, and reaction pathways, plus a perspective on functional dynamics and overall reaction rate chemistry.

3D representation

Figure 3. 19F NMR spectrum of defluorinase enzyme showing all nine fluoro-Trp resonances as a function of enzyme. Two residues in the active site are directly affected as product is formed. Two residues exhibit allostery from the addition of substrate under saturating conditions. The crystal structure of the substrate saturated state of the enzyme dimer is shown at right, with the Trp residues highlighted in green.

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