Article | Published:

Global metabolic inhibitors of sialyl- and fucosyltransferases remodel the glycome

Nature Chemical Biology volume 8, pages 661668 (2012) | Download Citation

Abstract

Despite the fundamental roles of sialyl- and fucosyltransferases in mammalian physiology, there are few pharmacological tools to manipulate their function in a cellular setting. Although fluorinated analogs of the donor substrates are well-established transition state inhibitors of these enzymes, they are not membrane permeable. By exploiting promiscuous monosaccharide salvage pathways, we show that fluorinated analogs of sialic acid and fucose can be taken up and metabolized to the desired donor substrate–based inhibitors inside the cell. Because of the existence of metabolic feedback loops, they also act to prevent the de novo synthesis of the natural substrates, resulting in a global, family-wide shutdown of sialyl- and/or fucosyltransferases and remodeling of cell-surface glycans. As an example of the functional consequences, the inhibitors substantially reduce expression of the sialylated and fucosylated ligand sialyl Lewis X on myeloid cells, resulting in loss of selectin binding and impaired leukocyte rolling.

  • Compound C14H20O9

    1,2,3,4-Tetra-O-acetyl-α,β-L-fucopyranose

  • Compound C12H17FO7

    2-Deoxy-2-fluoro-1,3,4-tri-O-acetyl-α,β-L-fucopyranose

  • Compound C14H19FO9

    6-Deoxy-6-fluoro-1,2,3,4-tetra-O-acetyl-α,β-L-galactopyranose

  • Compound C16H25N5O15P2

    Guanosine 5'-diphospho-β-L-fucopyranose

  • Compound C16H24FN5O14P2

    Guanosine 5'-diphospho-2-deoxy-2-fluoro-β-L-fucopyranose

  • Compound C16H24FN5O15P2

    Guanosine 5'-diphospho-6-deoxy-6-fluoro-β-L-galactopyranose

  • Compound C22H31NO14

    Methyl 5-acetamido-2,4,7,8,9-penta-O-acetyl-3,5-dideoxy-D-glycero-β-D-galacto-2-nonulopyranosate

  • Compound C22H30FNO14

    Methyl 5-acetamido-2,4,7,8,9-penta-O-acetyl-3,5-dideoxy-3-fluoro-D-erythro-β-L-manno-2-nonulopyranosate

  • Compound C22H30FNO14

    Methyl 5-acetamido-2,4,7,8,9-penta-O-acetyl-3,5-dideoxy-3-fluoro-D-erythro-β-L-gluco-2-nonulopyranosate

  • Compound C20H31N4O16P

    Cytidine-5'-monophospho 5-acetamido-3,5-dideoxy-D-glycero-β-D-galacto-2-nonulopyranosonic acid

  • Compound C20H30FN4O16P

    Cytidine-5'-monophospho 5-acetamido-3,5-dideoxy-3-fluoro-D-erythro-α-L-manno-2-nonulopyranosonic acid

  • Compound C20H30FN4O16P

    Cytidine-5'-monophospho 5-acetamido-3,5-dideoxy-3-fluoro-D-erythro-α-L-gluco-2-nonulopyranosonic acid

  • Compound C37H40N2O17

    Fluorescein-5-carbamidoethyl (β-D-galactopyranosyl)-(1->4)-2-acetamido-2-deoxy-β-D-glucopyranoside

  • Compound C48H57N3O25

    Fluorescein-5-carbamidoethyl (5-acetamido-3,5-dideoxy-D-glycero-α-D-galacto-2-nonulopyranosylonic acid)-(2->3)-(β-D-galactopyranosyl)-(1->4)-2-acetamido-2-deoxy-β-D-glucopyranoside

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Acknowledgements

This work was supported by US National Institutes of Health grants to J.C.P. (R01AI050143 and P01HL107151), C.D.R. (T32AI007606), K.L. (R01HL111969), C.T.L. (T32AI060536) and the Complex Carbohydrate Research Center (1 P41 RR018502-01) as well as by funding from the Biotechnology and Biological Sciences Research Council to A.D. and S.M.H. (BBF0083091).

Author information

Affiliations

  1. Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA.

    • Cory D Rillahan
    •  & James C Paulson
  2. Division of Molecular Biosciences, Faculty of Natural Sciences, Imperial College–London, London, UK.

    • Aristotelis Antonopoulos
    • , Anne Dell
    •  & Stuart M Haslam
  3. La Jolla Institute for Allergy and Immunology, Division of Inflammation Biology, La Jolla, California, USA.

    • Craig T Lefort
    •  & Klaus Ley
  4. Complex Carbohydrate Research Center, The University of Georgia, Athens, Georgia, USA.

    • Roberto Sonon
    •  & Parastoo Azadi

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Contributions

C.D.R. conceived of the idea, synthesized the inhibitors, designed the experiments and performed biochemical assays. A.A. performed the N- and O-linked glycan MS analysis. C.T.L. performed the rolling assays. R.S. performed the nucleotide sugar analysis. J.C.P., S.M.H., A.D., K.L. and P.A. supervised the research. C.D.R and J.C.P. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to James C Paulson.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Methods and Supplementary Results

Videos

  1. 1.

    Supplementary Video 1

    Rolling Velocity Measurement of control (DMSO) treated HL-60 cells on E-Selectin at 3 dynes/cm2

  2. 2.

    Supplementary Video 2

    Rolling Velocity Measurement of 2F-Fuc (2) treated HL-60 cells on ESelectin at 3 dynes/cm2

  3. 3.

    Supplementary Video 3

    Rolling Velocity Measurement of 3Fax-Neu5Ac (8) treated HL-60 cells on E-Selectin at 3 dynes/cm2

  4. 4.

    Supplementary Video 4

    Rolling Velocity Measurement of control (DMSO) treated HL-60 cells on P-Selectin at 3 dynes/cm2

  5. 5.

    Supplementary Video 5

    Rolling Velocity Measurement of 2F-Fuc (2) treated HL-60 cells on PSelectin at 3 dynes/cm2

  6. 6.

    Supplementary Video 6

    Rolling Velocity Measurement of 3Fax-Neu5Ac (8) treated HL-60 cells on P-Selectin at 3 dynes/cm2

  7. 7.

    Supplementary Video 7

    Tethering Analysis of Control (DMSO) treated HL-60 cells on E-Selectin

  8. 8.

    Supplementary Video 8

    Tethering Analysis of 2F-Fuc (2) treated HL-60 cells on E-Selectin

  9. 9.

    Supplementary Video 9

    Tethering Analysis of 3Fax-Neu5Ac (8) treated HL-60 cells on E-Selectin

  10. 10.

    Supplementary Video 10

    Tethering Analysis of Control (DMSO) treated HL-60 cells on PSelectin

  11. 11.

    Supplementary Video 11

    Tethering Analysis of 2F-Fuc (2) treated HL-60 cells on P-Selectin

  12. 12.

    Supplementary Video 12

    Tethering Analysis of 3Fax-Neu5Ac (8) treated HL-60 cells on PSelectin

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DOI

https://doi.org/10.1038/nchembio.999

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