WZB117 (2-Fluoro-6-(m-hydroxybenzoyloxy) Phenyl m-Hydroxybenzoate) Inhibits GLUT1-mediated Sugar Transport by Binding Reversibly at the Exofacial Sugar Binding Site

J Biol Chem. 2016 Dec 23;291(52):26762-26772. doi: 10.1074/jbc.M116.759175. Epub 2016 Nov 11.


WZB117 (2-fluoro-6-(m-hydroxybenzoyloxy) phenyl m-hydroxybenzoate) inhibits passive sugar transport in human erythrocytes and cancer cell lines and, by limiting glycolysis, inhibits tumor growth in mice. This study explores how WZB117 inhibits the erythrocyte sugar transporter glucose transport protein 1 (GLUT1) and examines the transporter isoform specificity of inhibition. WZB117 reversibly and competitively inhibits erythrocyte 3-O-methylglucose (3MG) uptake with Ki(app) = 6 μm but is a noncompetitive inhibitor of sugar exit. Cytochalasin B (CB) is a reversible, noncompetitive inhibitor of 3MG uptake with Ki(app) = 0.3 μm but is a competitive inhibitor of sugar exit indicating that WZB117 and CB bind at exofacial and endofacial sugar binding sites, respectively. WZB117 inhibition of GLUTs expressed in HEK293 cells follows the order of potency: insulin-regulated GLUT4 ≫ GLUT1 ≈ neuronal GLUT3. This may explain WZB117-induced murine lipodystrophy. Molecular docking suggests the following. 1) The WZB117 binding envelopes of exofacial GLUT1 and GLUT4 conformers differ significantly. 2) GLUT1 and GLUT4 exofacial conformers present multiple, adjacent glucose binding sites that overlap with WZB117 binding envelopes. 3) The GLUT1 exofacial conformer lacks a CB binding site. 4) The inward GLUT1 conformer presents overlapping endofacial WZB117, d-glucose, and CB binding envelopes. Interrogating the GLUT1 mechanism using WZB117 reveals that subsaturating WZB117 and CB stimulate erythrocyte 3MG uptake. Extracellular WZB117 does not affect CB binding to GLUT1, but intracellular WZB117 inhibits CB binding. These findings are incompatible with the alternating conformer carrier for glucose transport but are consistent with either a multisubunit, allosteric transporter, or a transporter in which each subunit presents multiple, interacting ligand binding sites.

Keywords: competitive inhibition; facilitated diffusion; glucose transport; ligand binding; ligand-binding protein; membrane transport; membrane transport protein; molecular docking; protein structure.

MeSH terms

  • 3-O-Methylglucose / metabolism*
  • Animals
  • Binding Sites
  • Biological Transport
  • Crystallography, X-Ray
  • Cytochalasin B / metabolism
  • Erythrocytes / drug effects
  • Erythrocytes / metabolism*
  • Glucose / metabolism*
  • Glucose Transporter Type 1 / chemistry
  • Glucose Transporter Type 1 / metabolism*
  • Glucose Transporter Type 3 / chemistry
  • Glucose Transporter Type 3 / metabolism
  • Glucose Transporter Type 4 / chemistry
  • Glucose Transporter Type 4 / metabolism
  • HEK293 Cells
  • Humans
  • Hydroxybenzoates / pharmacology*
  • Kinetics
  • Mice
  • Molecular Docking Simulation
  • Protein Conformation


  • Glucose Transporter Type 1
  • Glucose Transporter Type 3
  • Glucose Transporter Type 4
  • Hydroxybenzoates
  • SLC2A1 protein, human
  • SLC2A3 protein, human
  • SLC2A4 protein, human
  • WZB117
  • 3-O-Methylglucose
  • Cytochalasin B
  • Glucose

Associated data

  • PDB/4ZWC
  • PDB/4PYP