Nature Communications (Sep 2024)

Synthesis and screening of a library of Lewisx deoxyfluoro-analogues reveals differential recognition by glycan-binding partners

  • Kristian Hollingsworth,
  • Antonio Di Maio,
  • Sarah-Jane Richards,
  • Jean-Baptiste Vendeville,
  • David E. Wheatley,
  • Claire E. Council,
  • Tessa Keenan,
  • Hélène Ledru,
  • Harriet Chidwick,
  • Kun Huang,
  • Fabio Parmeggiani,
  • Andrea Marchesi,
  • Wengang Chai,
  • Ryan McBerney,
  • Tomasz P. Kamiński,
  • Matthew R. Balmforth,
  • Alexandra Tamasanu,
  • James D. Finnigan,
  • Carl Young,
  • Stuart L. Warriner,
  • Michael E. Webb,
  • Martin A. Fascione,
  • Sabine Flitsch,
  • M. Carmen Galan,
  • Ten Feizi,
  • Matthew I. Gibson,
  • Yan Liu,
  • W. Bruce Turnbull,
  • Bruno Linclau

DOI
https://doi.org/10.1038/s41467-024-51081-7
Journal volume & issue
Vol. 15, no. 1
pp. 1 – 14

Abstract

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Abstract Glycan-mediated interactions play a crucial role in biology and medicine, influencing signalling, immune responses, and disease pathogenesis. However, the use of glycans in biosensing and diagnostics is limited by cross-reactivity, as certain glycan motifs can be recognised by multiple biologically distinct protein receptors. To address this specificity challenge, we report the enzymatic synthesis of a 150-member library of site-specifically fluorinated Lewisx analogues (‘glycofluoroforms’) using naturally occurring enzymes and fluorinated monosaccharides. Subsequent incorporation of a subset of these glycans into nanoparticles or a microarray revealed a striking spectrum of distinct binding intensities across different proteins that recognise Lewisx. Notably, we show that for two proteins with unique binding sites for Lewisx, glycofluoroforms exhibited enhanced binding to one protein, whilst reduced binding to the other, with selectivity governed by fluorination patterns. We finally showcase the potential diagnostic utility of this approach in glycofluoroform-mediated bacterial toxin detection by lateral flow.