Molecular Gating of an Engineered Enzyme Captured in Real Time

Authors

Kokkonen, P., Sykora, J., Prokop, Z., Ghose, A., Bednar, D., Amaro, M., Beerens, K., Bidmanova, S., Slanska, M., Brezovsky, J., Damborsky, J., Hof, M.

Source

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 140: 17999–18008 (2018)

Abstract

Engineering dynamical molecular gates represents a widely applicable strategy for designing efficient biocatalysts. Here we analyzed the dynamics of a molecular gate artificially introduced into an access tunnel of the most efficient haloalkane dehalogenase using pre-steady-state kinetics, a single-molecule fluorescence spectroscopy and molecular dynamics. Photoinduced electron-transfer – fluorescence correlation spectroscopy (PET-FCS) has enabled real-time observation of molecular gating at single molecule level with the rate constants (kon = 1822 s-1, koff = 60 s-1) corresponding well with those from the pre-steady-state kinetics (k-1 = 1100 s-1, k1 = 20 s-1).

Full text

Citation

Kokkonen, P., Sykora, J., Prokop, Z., Ghose, A., Bednar, D., Amaro, M., Beerens, K., Bidmanova, S., Slanska, M., Brezovsky, J., Damborsky, J., Hof, M., 2018: Molecular Gating of an Engineered Enzyme Captured in Real Time. Journal of the American Chemical Society 140: 17999–18008.

Open Position: Laboratory Technician
Martin’s and Hana’s Success in Secondary-School Professional Activities
SoluProt: New Look
Michal Awarded for Excellence in Science
Czech Learned Society Award for Hana
Postdoc in Molecular Modeling Needed
Prof. Prokop at CCBIO Symposium on Industrial Biocatalysis