PDBsum entry 5foe

Transferase

PDB id: 5foe

Contents

Protein chains

429 a.a.

Ligands

NAG ×2

GDP ×2

EDO ×10

Waters ×531

References listed in PDB file

Key reference

• Title: A proactive role of water molecules in acceptor recognition by protein o-Fucosyltransferase 2.
• Authors: J.Valero-González, C.Leonhard-Melief, E.Lira-Navarrete, G.Jiménez-Osés, C.Hernández-Ruiz, M.C.Pallarés, I.Yruela, D.Vasudevan, A.Lostao, F.Corzana, H.Takeuchi, R.S.Haltiwanger, R.Hurtado-Guerrero.
• Ref.: Nat Chem Biol, 2016, 12, 240-246. [DOI no: 10.1038/nchembio.2019]
• PubMed id: 26854667

Abstract

Protein O-fucosyltransferase 2 (POFUT2) is an essential enzyme that fucosylates serine and threonine residues of folded thrombospondin type 1 repeats (TSRs). To date, the mechanism by which this enzyme recognizes very dissimilar TSRs has been unclear. By engineering a fusion protein, we report the crystal structure of Caenorhabditis elegans POFUT2 (CePOFUT2) in complex with GDP and human TSR1 that suggests an inverting mechanism for fucose transfer assisted by a catalytic base and shows that nearly half of the TSR1 is embraced by CePOFUT2. A small number of direct interactions and a large network of water molecules maintain the complex. Site-directed mutagenesis demonstrates that POFUT2 fucosylates threonine preferentially over serine and relies on folded TSRs containing the minimal consensus sequence C-X-X-S/T-C. Crystallographic and mutagenesis data, together with atomic-level simulations, uncover a binding mechanism by which POFUT2 promiscuously recognizes the structural fingerprint of poorly homologous TSRs through a dynamic network of water-mediated interactions.