Letter | Published:

Structural basis for glycosphingolipid transfer specificity

Nature volume 430, pages 10481053 (26 August 2004) | Download Citation



Lipid transfer proteins are important in membrane vesicle biogenesis and trafficking, signal transduction and immunological presentation processes1,2,3. The conserved and ubiquitous mammalian glycolipid transfer proteins (GLTPs) serve as potential regulators of cell processes mediated by glycosphingolipids, ranging from differentiation and proliferation to invasive adhesion, neurodegeneration and apoptosis4,5. Here we report crystal structures of apo-GLTP (1.65 Å resolution) and lactosylceramide-bound (1.95 Å) GLTP, in which the bound glycosphingolipid is sandwiched, after adaptive recognition, within a previously unknown two-layer all-α-helical topology. Glycosphingolipid binding specificity is achieved through recognition and anchoring of the sugar-amide headgroup to the GLTP recognition centre by hydrogen bond networks and hydrophobic contacts, and encapsulation of both lipid chains, in a precisely oriented manner within a ‘moulded-to-fit’ hydrophobic tunnel. A cleft-like conformational gating mechanism, involving two interhelical loops and one α-helix of GLTP, could enable the glycolipid chains to enter and leave the tunnel in the membrane-associated state. Mutation and functional analyses of residues in the glycolipid recognition centre and within the hydrophobic tunnel support a framework for understanding how GLTPs acquire and release glycosphingolipids during lipid intermembrane transfer and presentation processes.

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We thank the personnel at SBC beamline 19BM of the Advanced Photon Source beamline staff for assistance with data collection from multiwavelength anomalous dispersion; A. Serganov for technical support; X. Lin, T. Chung and H. Pike for their contributions to the cloning and expression of the recombinant human GLTP; X.-M. Li for synthesizing and purifying N-18:1 lactosylceramide; A. J. Windebank for help with DNA sequencing at the Mayo Molecular Biology Core Facility; T. Burghardt for help with recording near-ultraviolet CD spectra; and S. Venyaminov in the Franklyn Prendergast laboratory for recording the far-ultraviolet CD spectra. This research was supported by NIH and the Hormel Foundation. Use of the ANL SBC beamlines at the APS was supported by the US Department of Energy, Office of Energy Research.

Author information

Author notes

    • Lucy Malinina
    •  & Margarita L. Malakhova

    These authors contributed equally to this work


  1. Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA

    • Lucy Malinina
    • , Alexei Teplov
    •  & Dinshaw J. Patel
  2. Hormel Institute, University of Minnesota, Austin, Minnesota 55912, USA

    • Margarita L. Malakhova
    •  & Rhoderick E. Brown


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Competing interests

The authors declare that they have no competing financial interests.

Corresponding authors

Correspondence to Rhoderick E. Brown or Dinshaw J. Patel.

Supplementary information

Rich text format

  1. 1.

    Supplementary Table S1

    X-ray data collection and refinement statistics.

PDF files

  1. 1.

    Supplementary Figure S1

    A two-layer topology of the γ-helices.

  2. 2.

    Supplementary Figure S2

    Structure of the lactosylceramide-GLTP complex.

  3. 3.

    Supplementary Figure S3

    Schematic showing the hydrogen bonding between lactosylceramide and protein side chains in the complex.

  4. 4.

    Supplementary Figure S4

    Glycolipid transfer activities of wtGLTP and representative GLTPs with point mutations.

  5. 5.

    Supplementary Figure S5

    Superposition of wild type GLTP and the D48V mutant.

  6. 6.

    Supplementary Figure S6

    Far UV CD spectra of GLTP mutants.

  7. 7.

    Supplementary Figure S7

    Near UV CD spectra of GLTP mutants.

  8. 8.

    Supplementary Figure S8

    Electron density map, sequence and secondary structure elements for apo-GLTP.

  9. 9.

    Supplementary Figure S9

    Omit electron density map for a ligand.

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