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Abstract

The glycans displayed on mammalian cells can differ markedly from those on microbes. Such differences could, in principle, be 'read' by carbohydrate-binding proteins, or lectins. We used glycan microarrays to show that human intelectin-1 (hIntL-1) does not bind known human glycan epitopes but does interact with multiple glycan epitopes found exclusively on microbes: β-linked D-galactofuranose (β-Galf), D-phosphoglycerol–modified glycans, heptoses, D-glycero-D-talo-oct-2-ulosonic acid (KO) and 3-deoxy-D-manno-oct-2-ulosonic acid (KDO). The 1.6-Å-resolution crystal structure of hIntL-1 complexed with β-Galf revealed that hIntL-1 uses a bound calcium ion to coordinate terminal exocyclic 1,2-diols. N-acetylneuraminic acid (Neu5Ac), a sialic acid widespread in human glycans, has an exocyclic 1,2-diol but does not bind hIntL-1, probably owing to unfavorable steric and electronic effects. hIntL-1 marks only Streptococcus pneumoniae serotypes that display surface glycans with terminal 1,2-diol groups. This ligand selectivity suggests that hIntL-1 functions in microbial surveillance.

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Acknowledgements

This research was supported by the US National Institutes of Health (NIH) (R01GM55984 and R01AI063596 (L.L.K.)). D.A.W. thanks the US National Science Foundation (NSF) and the NIH Chemistry-Biology Interface Training Program (T32 GM008505) for fellowships. K.W. was supported by a fellowship from the Development and Promotion of Science and Technology Talents Project of Thailand. M.B.K. and H.L.H. were supported by the NIH (F32 GM100729 to M.B.K. and T32 GM008505 to H.L.H.). L.C.Z. was supported by a UW–Madison Hilldale Fellowship. R.A.S. thanks the American Chemical Society Division of Medicinal Chemistry for a fellowship. The glycan array experiments were made possible by the Consortium for Functional Glycomics (NIH NIGMS GM062116 and GM98791 (J.C.P.)), which supported the Glycan Array Synthesis Core at The Scripps Research Institute and the Protein-Glycan Interaction Resource (Emory University School of Medicine). These resources assisted with analysis of samples on the array. Printing and processing the furanoside array was supported through the US National Center for Functional Glycomics supported by NIH NIGMS (P41GM103694 (R.D.C.)). SPR experiments were performed at the University of Wisconsin (UW)−Madison Biophysics Instrumentation Facility, which is supported by UW–Madison, NSF grant BIR-9512577 and NIH grant S10 RR13790. Flow cytometry data were obtained at the UW–Madison Carbone Cancer Center (P30 CA014520), and microscopy images were acquired at the UW−Madison W.M. Keck Laboratory for Biological Imaging (1S10RR024715). The UW−Madison Chemistry NMR facility is supported by the NSF (CHE-9208463 and CHE-9629688) and NIH (1s10 RR08389). Use of the Advanced Photon Source at the Argonne National Laboratory was supported by the US Department of Energy (contract DE-AC02-06CH11357), and the LS-CAT Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (grant 085P1000817). We thank J.M. Fishman for assistance in preparing the synthetic methods and M.R. Levengood, A.H. Courtney and D.R. McCaslin for thoughtful discussions. We thank M.R. Richards (University of Alberta) for helpful discussions.

Author information

Author notes

    • Matthew B Kraft
    •  & Rebecca A Splain

    Present Addresses: Gilead Sciences, Foster City, California, USA (M.B.K.) and Global API Chemistry, GlaxoSmithKline, King of Prussia, Pennsylvania, USA (R.A.S.).

Affiliations

  1. Department of Biochemistry, University of Wisconsin−Madison, Madison, Wisconsin, USA.

    • Darryl A Wesener
    • , Lucas C Zarling
    •  & Laura L Kiessling
  2. Department of Chemistry, University of Wisconsin−Madison, Madison, Wisconsin, USA.

    • Kittikhun Wangkanont
    • , Matthew B Kraft
    • , Heather L Hodges
    • , Rebecca A Splain
    •  & Laura L Kiessling
  3. Department of Cell and Molecular Biology, Scripps Research Institute, La Jolla, California, USA.

    • Ryan McBride
    •  & James C Paulson
  4. Department of Chemical Physiology, Scripps Research Institute, La Jolla, California, USA.

    • Ryan McBride
    •  & James C Paulson
  5. Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.

    • Xuezheng Song
    • , David F Smith
    •  & Richard D Cummings
  6. Glycomics Center, Emory University School of Medicine, Atlanta, Georgia, USA.

    • Xuezheng Song
    • , David F Smith
    •  & Richard D Cummings
  7. Department of Bacteriology, University of Wisconsin−Madison, Madison, Wisconsin, USA.

    • Katrina T Forest

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Contributions

D.A.W. and L.L.K. conceived the project. D.A.W., K.W. and L.L.K. planned the experiments, analyzed the data and wrote the paper, with input from all the other authors. Cloning, protein expression and biochemical experiments were performed by D.A.W. and L.C.Z. Microscopy was performed by H.L.H. Baculovirus was made by K.W. The carbohydrate ligands were synthesized and characterized by M.B.K. and R.A.S. The furanoside glycan microarray was constructed and analyzed with the mammalian glycan microarray by X.S., D.F.S. and R.D.C. The microbial glycan array was constructed and analyzed by R.M. and J.C.P. Protein crystallization and structure determination were performed by K.W. and K.T.F.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Laura L Kiessling.

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    Supplementary Text and Figures

    Supplementary Figures 1–7 and Supplementary Note

Excel files

  1. 1.

    Supplementary Table 1

    Human IntL-1 binding to furanoside glycan array

  2. 2.

    Supplementary Table 2

    Human IntL-1 binding to mammalian glycan array

  3. 3.

    Supplementary Table 3

    Human IntL-1 binding to microbial glycan array

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DOI

https://doi.org/10.1038/nsmb.3053

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