Fluorescence spectroscopy as a tool to investigate protein interactions

Translational and rotational motions of proteins in a protein crowded environment

“…It is therefore important to incorporate crowding effects in the design and interpretation of in vitro studies [9,10], in order appropriately to understand and quantify how kinetic properties of specific macromolecules in physiological media are modified by the presence of other, neutral (non-interacting) macromolecules in their neighborhood. Among other biophysical techniques, methods based on fluorescence spectroscopy [11][12][13] appear to be very convenient for interaction studies in crowded media due to the possibility of specifically labeling only the tracer protein with extrinsic fluorescent dyes, which in this way can be distinguished easily from the crowding macromolecules. These techniques allow characterization and quantification of the hydrodynamic properties of free and bound species, both in crowded model solutions [14,15] and in whole living cells [6,[16][17][18], using their fluorescence.…”

Structural Dynamics of C-domain of Cardiac Troponin I Protein in Reconstituted Thin Filament

“…Fluorescence anisotropy is then defined by the relative fluorescence intensity of the vertically polarized emission with respect to the intensity of the total emission (19). In principle, the steady-state fluorescence anisotropy (r) of the fluorophore-labeled protein can be approximated using Perrin's equation (20,21),…”

Fundamental Chemical and Structural Principles

“…Fluorescence spectroscopy [181] is widely used in peptide and protein chemistry, either observing intrinsic fluorophors (Trp, Tyr), fluorescent cofactors, or extrinsic fluorophors that are used to label the protein. As fluorescence spectroscopy involves electronic transitions, it can be applied to study very fast processes.…”