title: The promiscuous protein binding ability of erythrosine B studied by metachromasy (metachromasia).
abstract: The present study aims to elucidate aspects of the protein binding ability of erythrosine B (ErB), a poly-iodinated xanthene dye and an FDA-approved food colorant (FD&C Red No. 3), which we have identified recently as a promiscuous inhibitor of protein-protein interactions (PPIs) with a remarkably consistent median inhibitory concentration (IC50 ) in the 5- to 30-μM range. Because ErB exhibits metachromasy, that is, color change upon binding to several proteins, we exploited this property to quantify its binding to proteins such as bovine serum albumin (BSA) and CD40L (CD154) and to determine the corresponding binding constants (Kd ) and stoichiometry (nb ) using spectrophotometric methods. Binding was reversible, and the estimated affinities for both protein targets obtained here (Kd values of 14 and 20 μM for BSA and CD40L, respectively) were in good agreement with that expected from the PPI inhibitory activity of ErB. A stoichiometry greater than one was observed both for CD40L and BSA binding (nb of 5-6 and 8-9 for BSA and CD40L, respectively), indicating the possibility of nonspecific binding of the flat and rigid ErB molecule at multiple sites, which could explain the promiscuous PPI inhibitory activity if some of these overlap with the binding site of the protein partner and interfere with the binding.
authors: Department of Molecular and Cellular Pharmacology and Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, FL, USA.
authors: Lakshmi
authors: L
authors: Ganesan
authors: Peter
authors: P
authors: Buchwald
mesh_terms: D000818:Animals; D023201:CD40 Ligand / Q000737:chemistry; D002417:Cattle; D004923:Erythrosine / Q000737:chemistry*; D005505:Food Coloring Agents / Q000737:chemistry*; D006801:Humans; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013057:Spectrum Analysis
chemical_list: D005505:Food Coloring Agents; D023201:CD40 Ligand; D012710:Serum Albumin, Bovine; D004923:Erythrosine