Structural basis of substrate recognition and catalysis by fucosyltransferase 8

Michael A. Järvå, Marija Dramicanin, James P. Lingford, Runyu Mao, Alan John, Kate E. Jarman, Rhys Grinter, Ethan D. Goddard-Borger

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

Fucosylation of the innermost GlcNAc of N-glycans by fucosyltransferase 8 (FUT8) is an important step in the maturation of complex and hybrid N-glycans. This simple modification can dramatically affect the activities and half-lives of glycoproteins, effects that are relevant to understanding the invasiveness of some cancers, development of mAb therapeutics, and the etiology of a congenital glycosylation disorder. The acceptor substrate preferences of FUT8 are well-characterized and provide a framework for understanding N-glycan maturation in the Golgi; however, the structural basis of these substrate preferences and the mechanism through which catalysis is achieved remain unknown. Here we describe several structures of mouse and human FUT8 in the apo state and in complex with GDP, a mimic of the donor substrate, and with a glycopeptide acceptor substrate at 1.80 –2.50 Å resolution. These structures provide insights into a unique conformational change associated with donor substrate binding, common strategies employed by fucosyltransferases to coordinate GDP, features that define acceptor substrate preferences, and a likely mechanism for enzyme catalysis. Together with molecular dynamics simulations, the structures also revealed how FUT8 dimerization plays an important role in defining the acceptor substrate-binding site. Collectively, this information significantly builds on our understanding of the core fucosylation process.

Original languageEnglish
Pages (from-to)6677-6688
Number of pages13
JournalJournal of Biological Chemistry
Volume295
Issue number19
DOIs
Publication statusPublished - 8 May 2020

Keywords

  • glycosyltransferase
  • N-linked glycosylation
  • fucosyltransferase
  • glycobiology
  • core fucose
  • enzyme mechanism
  • structural biology
  • fucosyltransferase 8 (FUT8)
  • N-acetylglucosamine (GlcNAc)
  • protein maturation
  • enzyme evolution

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