Ligand-binding assay based on microfluidic chemotaxis of porphyrin receptors.

Recent studies have shown that enzymes undergo chemotaxis up substrate gradients during catalysis. One important avenue to identify the molecular level origins of this phenomenon is the ligand-protein binding that occurs even in the absence of catalytic turnover. Here, the chemotaxis of zinc porphyrin as a cofactor mimic was observed by imposing a concentration gradient of organic amines in the microfluidic device. Their axial ligations led to the directed motions of porphyrin receptors. The dissociation constant for selected recognition could be obtained by measuring the chemotactic shift as a function of ligand content, which is associated with both the binding strength and the steric hindrance of the specific ligand. Finally, a statistical thermodynamic model was derived, relating the change of Gibbs free energy (Δ G ) in the binding process to the directional migration of receptors. The theoretical model agreed quantitatively with experimental results, elucidating that Δ G of reversible binding essentially drives molecular chemotaxis.