title: Molecular basis of ligand recognition and transport by glucose transporters.
abstract: The major facilitator superfamily glucose transporters, exemplified by human GLUT1-4, have been central to the study of solute transport. Using lipidic cubic phase crystallization and microfocus X-ray diffraction, we determined the structure of human GLUT3 in complex with D-glucose at 1.5 Å resolution in an outward-occluded conformation. The high-resolution structure allows discrimination of both α- and β-anomers of D-glucose. Two additional structures of GLUT3 bound to the exofacial inhibitor maltose were obtained at 2.6 Å in the outward-open and 2.4 Å in the outward-occluded states. In all three structures, the ligands are predominantly coordinated by polar residues from the carboxy terminal domain. Conformational transition from outward-open to outward-occluded entails a prominent local rearrangement of the extracellular part of transmembrane segment TM7. Comparison of the outward-facing GLUT3 structures with the inward-open GLUT1 provides insights into the alternating access cycle for GLUTs, whereby the C-terminal domain provides the primary substrate-binding site and the amino-terminal domain undergoes rigid-body rotation with respect to the C-terminal domain. Our studies provide an important framework for the mechanistic and kinetic understanding of GLUTs and shed light on structure-guided ligand design.
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.|Tsinghua-Peking Center for Life Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.|Tsinghua-Peking Center for Life Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.|Tsinghua-Peking Center for Life Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Chuangye
authors.initials: C
authors.lastname: Yan
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.|Tsinghua-Peking Center for Life Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Ke
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Tsinghua-Peking Center for Life Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xiong
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.
authors.forename: Wenlin
authors.initials: W
authors.lastname: Ren
authors.affiliation: Advanced Photon Technology Division, Research Infrastructure Group, SR Life Science Instrumentation Unit, RIKEN/SPring-8 Center, 1-1-1 Kouto Sayo-cho Sayo-gun, Hyogo 679-5148 Japan.|Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Kunio
authors.initials: K
authors.lastname: Hirata
authors.affiliation: Advanced Photon Technology Division, Research Infrastructure Group, SR Life Science Instrumentation Unit, RIKEN/SPring-8 Center, 1-1-1 Kouto Sayo-cho Sayo-gun, Hyogo 679-5148 Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: Center for Structural Biology, Tsinghua University, Beijing 100084, China.
authors.forename: Shilong
authors.initials: S
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Membrane Biology, Tsinghua University, Beijing 100084, China.|Center for Structural Biology, Tsinghua University, Beijing 100084, China.|Tsinghua-Peking Center for Life Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Nieng
authors.initials: N
authors.lastname: Yan
mesh_terms: D001665:Binding Sites; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D018360:Crystallography, X-Ray; D005947:Glucose / Q000737:chemistry* / Q000378:metabolism*; D051272:Glucose Transporter Type 1 / Q000737:chemistry / Q000378:metabolism; D051274:Glucose Transporter Type 3 / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008320:Maltose / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012399:Rotation; D013329:Structure-Activity Relationship
chemical_list: D051272:Glucose Transporter Type 1; D051274:Glucose Transporter Type 3; D008024:Ligands; D011994:Recombinant Proteins; C495677:SLC2A1 protein, human; C495687:SLC2A3 protein, human; D008320:Maltose; D005947:Glucose