Time-Dependent Fluorescence Spectroscopy to Quantify Complex Binding Interactions.

ABSTRACT: Measuring the binding affinity for proteins that can aggregate or undergo complex binding motifs presents a variety of challenges. In this study, fluorescence lifetime measurements using intrinsic tryptophan fluorescence were performed to address these challenges and to quantify the binding of a series of carbohydrates and carbohydrate-functionalized dendrimers to recombinant human galectin-3. Collectively, galectins represent an important target for study; in particular, galectin-3 plays a variety of roles in cancer biology. Galectin-3 binding dissociation constants (K D) were quantified: lactoside (73 ± 4 ?M), methyllactoside (54 ± 10 ?M), and lactoside-functionalized G(2), G(4), and G(6)-PAMAM dendrimers (120 ± 58 ?M, 100 ± 45 ?M, and 130 ± 25 ?M, respectively). The chosen examples showcase the widespread utility of time-dependent fluorescence spectroscopy for determining binding constants, including interactions for which standard methods have significant limitations.

SUBMITTER: Bernhard SP 

PROVIDER: S-EPMC7675582 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature