Chapter 2 - Fluorescence-based techniques to assess biomolecular structure and dynamics

Abstract

Fluorescence is a phenomenon of broad appeal and several applications. Among the most important ones are fluorescence-based assays and methods that allow the analysis of biomolecular structure and dynamics. For structural studies, one can assess the relative orientation of protein fragments to the polarity of the environment and describe conformational changes. Moreover, both small ligand binding and oligomerization are easily studied with fluorescence-based techniques. Dissociation constant and other kinetics parameters can also be calculated from such data. Finally, biomolecules might also be identified and tracked in vivo in real time and at high resolution. The biomolecule of interest may be autofluorescent, or it may need external labeling; the availability of different fluorophores is also discussed. Here, we discuss essential applications of fluorescence-based methods providing an overview of the vast possibilities they offer for researchers.