title: Quantitative Thermodynamic Analyses of Spectroscopic Titration Curves.
abstract: Elucidation of ligand - macromolecule interactions requires detailed knowledge of energetics of the formed complexes. Spectroscopic methods are most commonly used in characterizing molecular interactions in solution. The methods do not require large quantities of material and most importantly, do not perturb the studied reactions. However, spectroscopic methods absolutely require the determination of the relationship between the observed signal and the degree of binding in order to obtain meaningful interaction parameters. In other words, the meaningful, thermodynamic interaction parameters can be only determined if the relationship between the observed signal and the degree of binding is determined and not assumed, based on anad hocmodel of the relationship. The approaches discussed here allow an experimenter to quantitatively determine the degree of binding and the free ligand concentration,i.e., they enable to construct thermodynamic binding isotherms in a model-independent fashion.
authors.affiliation: Department of Obstetrics and Gynecology, The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, Texas 77555-1053.
authors.forename: Wlodzimierz
authors.initials: W
authors.lastname: Bujalowski
authors.affiliation: Department of Biochemistry and Molecular Biology, The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, Texas 77555-1053.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Jezewska
keywords: Binding Isotherms; Fluorescence; Spectroscopic Titrations; Thermodynamics