7-Hydroxycoumarins Are Affinity-Based Fluorescent Probes for Competitive Binding Studies of Macrophage Migration Inhibitory Factor
Zhangping Xiao, Deng Chen, Shanshan Song, Ramon van der Vlag, Petra E. van der Wouden, Ronald van Merkerk, Robbert H. Cool, Anna K. H. Hirsch, Barbro N. Melgert, Wim J. Quax, Gerrit J. Poelarends, Frank J. Dekker
Abstract
= 18 ± 1 nM) that binds with concomitant quenching of its fluorescence. This property enabled development of a novel competition-based assay format to quantify MIF binding. We also demonstrated that the 7-hydroxycoumarin fluorophore interfered with the MIF-CD74 interaction and inhibited proliferation of A549 cells. Thus, we provide a high-affinity MIF binder as a novel tool to advance MIF-oriented research.
Topics & Concepts
Macrophage migration inhibitory factorChemistryFluorophoreBiochemistryEnzymeStructure–activity relationshipCytokineFluorescenceIn vitroBiologyImmunologyPhysicsQuantum mechanicsMacrophage Migration Inhibitory Factor