FUT10 and FUT11 are protein O-fucosyltransferases that modify protein EMI domains

Huilin Hao; Youxi Yuan; Atsuko Ito; Benjamin M Eberand; Harry Tjondro; Michelle Cielesh; Nicholas Norris; Cesar L Moreno; Joshua W C Maxwell; G Gregory Neely; Richard J Payne; Melkam A Kebede; Ramona J Bieber Urbauer; Freda H Passam; Mark Larance; Robert S Haltiwanger

doi:10.1038/s41589-024-01815-x

FUT10 and FUT11 are protein O-fucosyltransferases that modify protein EMI domains

Nat Chem Biol. 2025 Jan 7. doi: 10.1038/s41589-024-01815-x. Online ahead of print.

Authors

Huilin Hao¹, Youxi Yuan¹, Atsuko Ito^{1

2}, Benjamin M Eberand³, Harry Tjondro⁴, Michelle Cielesh³, Nicholas Norris³, Cesar L Moreno⁵, Joshua W C Maxwell^{6

7}, G Gregory Neely⁵, Richard J Payne^{6

7}, Melkam A Kebede³, Ramona J Bieber Urbauer¹, Freda H Passam⁴, Mark Larance⁸, Robert S Haltiwanger⁹

Affiliations

¹ Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.
² Regional Fish Institute, Ltd., Kyoto, Japan.
³ Charles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, New South Wales, Australia.
⁴ Central Clinical School, The University of Sydney, Sydney, New South Wales, Australia.
⁵ Charles Perkins Centre, School of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, New South Wales, Australia.
⁶ School of Chemistry, Faculty of Science, The University of Sydney, Sydney, New South Wales, Australia.
⁷ Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Sydney, Sydney, New South Wales, Australia.
⁸ Charles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, New South Wales, Australia. [email protected].
⁹ Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA. [email protected].

PMID: 39775168
DOI: 10.1038/s41589-024-01815-x

Abstract

O-Fucosylation plays crucial roles in various essential biological events. Alongside the well-established O-fucosylation of epidermal growth factor-like repeats by protein O-fucosyltransferase 1 (POFUT1) and thrombospondin type 1 repeats by POFUT2, we recently identified a type of O-fucosylation on the elastin microfibril interface (EMI) domain of Multimerin-1 (MMRN1). Here, using AlphaFold2 screens, co-immunoprecipitation, enzymatic assays combined with mass spectrometric analysis and CRISPR-Cas9 knockouts, we demonstrate that FUT10 and FUT11, originally annotated in UniProt as α1,3-fucosyltransferases, are actually POFUTs responsible for modifying EMI domains; thus, we renamed them as POFUT3 and POFUT4, respectively. Like POFUT1/2, POFUT3/4 function in the endoplasmic reticulum, require folded domain structures for modification and participate in a non-canonical endoplasmic reticulum quality control pathway for EMI domain-containing protein secretion. This finding expands the O-fucosylation repertoire and provides an entry point for further exploration in this emerging field of O-fucosylation.

Abstract

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