Recombinant receptor-binding domain of diphtheria toxin increases the potency of curcumin by enhancing cellular uptake

Mol Pharm. 2014 Jan 6;11(1):208-17. doi: 10.1021/mp400378x. Epub 2013 Nov 22.

Abstract

Diphtheria toxin (DT) binds to a specific cell surface receptor, gets internalized, and causes cytotoxicity through its catalytic domain. The toxicity of DT is used in several therapeutic molecules. Here, we have exploited the receptor-binding ability of DT to increase cellular uptake of curcumin, a hydrophobic molecule with low bioavailability and cellular uptake. We have expressed only the receptor-binding domain of DT (RDT) in Escherichia coli. Purified RDT binds to the receptor with an affinity equivalent to that of full-length DT. It also binds to curcumin forming a curcumin-RDT complex, and this increases the fluorescence intensity and fluorescence lifetime of curcumin. The curcumin-RDT complex binds to the receptor and associates with human glioblastoma cells (U-87 MG) expressing the receptor. The cellular uptake of curcumin is higher for the curcumin-RDT complex than curcumin alone. This increase in uptake enhances the antiproliferative effect of curcumin and induces apoptosis of these cells even at a lower dose.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Cell Survival / drug effects
  • Chromatography, High Pressure Liquid
  • Curcumin / pharmacology*
  • Diphtheria Toxin / metabolism*
  • Drug Carriers
  • Flow Cytometry
  • Glioblastoma / drug therapy*
  • Glioblastoma / metabolism
  • Glioblastoma / pathology
  • Humans
  • Microscopy, Fluorescence
  • Protein Binding
  • Protein Conformation
  • Receptors, Cell Surface / metabolism*
  • Recombinant Proteins / metabolism*
  • Surface Plasmon Resonance
  • Tumor Cells, Cultured

Substances

  • Antineoplastic Agents
  • Diphtheria Toxin
  • Drug Carriers
  • Receptors, Cell Surface
  • Recombinant Proteins
  • Curcumin