Dynamics and Hydration of Proteins Viewed by Fluorescence Methods: Investigations for Protein Engineering and Synthetic Biology
Authors
Sykora, J., Prokop, Z., Damborsky, J., Abhinav, Hof, M., Amaro, M.
Book
FLUORESCENCE SPECTROSCOPY AND MICROSCOPY IN BIOLOGY: 509–532
Editor
Šachl, R., Amaro, M. (Eds.)
Publisher
Springer, Cham
Abstract
Protein engineering and synthetic biology are currently very active areas of research and development. In the pursuit of engineering proteins with specific capabilities, it has become evident that the scrutiny of structural and geometrical properties does not suffice to achieve the proposed goals. The dynamics and hydration of specific protein areas seem to be of higher influence than it has been once thought. This chapter introduces three different fluorescence spectroscopy techniques (time-dependent fluorescent shift, HMC hydration assay based on unnatural amino acid fluorescence, and photoinduced electron transfer–fluorescence correlation spectroscopy) that allow for assessing the dynamics and hydration of proteins in a site-specific fashion and showcase their usefulness in advancing the design of more efficient enzymes. Systematic application of these techniques to various biomolecular systems will allow a thorough description of these important protein properties, which are rarely taken into account during protein engineering and synthetic biology projects.