Crystallization and structural determination of the human glucose transporters GLUT1 and GLUT3

Dong Deng, Nieng Yan

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages15-29
Number of pages15
DOIs
StatePublished - Jan 1 2018
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1713
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Crystallization
  • GLUT1
  • GLUT3
  • Glucose transporters
  • Glut
  • Protein purification

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  • Cite this

    Deng, D., & Yan, N. (2018). Crystallization and structural determination of the human glucose transporters GLUT1 and GLUT3. In Methods in Molecular Biology (pp. 15-29). (Methods in Molecular Biology; Vol. 1713). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7507-5_2