Molecules (Apr 2021)

Protein <i>O</i>-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution

  • Erandi Lira-Navarrete,
  • María Carmen Pallarés,
  • Fabio Castello,
  • Maria J. Ruedas-Rama,
  • Angel Orte,
  • Anabel Lostao,
  • Ramón Hurtado-Guerrero

DOI
https://doi.org/10.3390/molecules26082105
Journal volume & issue
Vol. 26, no. 8
p. 2105

Abstract

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Protein O-fucosyltransferase 1 (PoFUT1) is a GT-B fold enzyme that fucosylates proteins containing EGF-like repeats. GT-B glycosyltransferases have shown a remarkable grade of plasticity adopting closed and open conformations as a way of tuning their catalytic cycle, a feature that has not been observed for PoFUT1. Here, we analyzed Caenorhabditis elegans PoFUT1 (CePoFUT1) conformational behavior in solution by atomic force microscopy (AFM) and single-molecule fluorescence resonance energy transfer (SMF-FRET). Our results show that this enzyme is very flexible and adopts mainly compact conformations and to a lesser extend a highly dynamic population that oscillates between compact and highly extended conformations. Overall, our experiments illustrate the inherent complexity of CePoFUT1 dynamics, which might play a role during its catalytic cycle.

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