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Predicting the Three-Dimensional Structure of the Human Facilitative Glucose Transporter Glut1 by a Novel Evolutionary Homology Strategy: Insights on the Molecular Mechanism of Substrate Migration, and Binding Sites for Glucose and Inhibitory Molecules

Alexis Salas-Burgos ^*, Pavel Iserovich ^*, Felipe Zuniga , Juan Carlos Vera  and Jorge Fischbarg ^* 

^* Department of Ophthalmology, College of Physicians & Surgeons, Columbia University, New York, New York; Department of Pathophysiology, School of Sciences, University of Concepción, Chile; and Department of Physiology and Biophysics, College of Physicians & Surgeons, Columbia University, New York, New York

Correspondence: Address reprint requests to Jorge Fischbarg, Dept. of Physiology, College of Physicians & Surgeons, 630 West 168th St., New York, NY 10032. Tel.: 212-305-9092; Fax: 212-305-2461; E-mail: jf20{at}columbia.edu.

The glucose transporters (GLUT/SLC2A) are members of the major facilitator superfamily. Here, we generated a three-dimensional model for Glut1 using a two-step strategy: 1), GlpT structure as an initial homology template and 2), evolutionary homology using glucose-6-phosphate translocase as a template. The resulting structure (PDB No. 1SUK) exhibits a water-filled passageway communicating the extracellular and intracellular domains, with a funnel-like exofacial vestibule (infundibulum), followed by a 15 Å-long x 8 Å-wide channel, and a horn-shaped endofacial vestibule. Most residues which, by mutagenesis, are crucial for transport delimit the channel, and putative sugar recognition motifs (QLS, QLG) border both ends of the channel. On the outside of the structure there are two positively charged cavities (one exofacial, one endofacial) delimited by ATP-binding Walker motifs, and an exofacial large side cavity of yet unknown function. Docking sites were found for the glucose substrate and its inhibitors: glucose, forskolin, and phloretin at the exofacial infundibulum; forskolin, and phloretin at an endofacial site next to the channel opening; and cytochalasin B at a positively charged endofacial pocket 3 Å away from the channel. Thus, 1SUK accounts for practically all biochemical and mutagenesis evidence, and provides clues for the transport process.

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