Protein O-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution Lira Navarrete, Erandi Castello, Fabio Ruedas Rama, María José Orte Gutiérrez, Ángel Glycosyltransferases O-fucosylation Protein dynamics Atomic force microscopy Singlemolecule methods 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. 2021-04-13T10:04:03Z 2021-04-13T10:04:03Z 2021 journal article Lira-Navarrete, E.; Pallarés, M.C.; Castello, F.; Ruedas-Rama, M.J.; Orte, A.; Lostao, A.; Hurtado-Guerrero, R. Protein O-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution. Molecules 2021, 26, 2105. https:// doi.org/10.3390/molecules26082105 http://hdl.handle.net/10481/67929 10.3390/molecules26082105 eng http://creativecommons.org/licenses/by/3.0/es/ open access Atribución 3.0 España MDPI