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Crystallization and Structural Determination of the Human Glucose Transporters GLUT1 and GLUT3

  • Dong Deng
  • Nieng Yan
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1713)

Abstract

Overexpression, purification, and crystallization of eukaryotic membrane proteins represent a major challenge for structural biology. In recent years, we have solved the crystal structures of the human glucose transporters GLUT1 in the inward-open conformation at 3.17 Å resolution and GLUT3 in the outward-open and occluded conformations at 2.4 and 1.5 Å resolutions, respectively. Structural elucidation of these transporters in three distinct functional states reveal the molecular basis for the alternating access transport cycle of this prototypal solute carrier family. It established the molecular foundation for future dynamic and kinetic investigations of these GLUTs, and will likely facilitate structure-based ligand development. In this chapter, we present the detailed protocols of recombinant protein expression, purification, and crystallization of GLUT1 and GLUT3, which may help the pursuit of structural elucidation of other eukaryotic membrane proteins.

Keywords

Glucose transporters Glut GLUT1 GLUT3 Protein purification Crystallization 

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Membrane BiologySchool of Life Sciences, Tsinghua UniversityBeijingChina
  2. 2.Beijing Advanced Innovation Center for Structural Biology, Tsinghua-Peking Center for Life SciencesSchool of Life Sciences and School of Medicine, Tsinghua UniversityBeijingChina
  3. 3.Key Laboratory of Birth Defects and Related Diseases of Women and Children of MOE, State Key Laboratory of BiotherapyWest China Second Hospital, Sichuan University and National Collaborative Innovation CenterChengduChina

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