8AY1

Crystal structure of the C. elegans POFUT2 (CePoFUT2) triple mutant (R298K-R299K-A418C) in complex with the Rattus norvegicus TSR4 single mutant (E10C) from F-spondin

Summary for 8AY1

• Entry DOI: 10.2210/pdb8ay1/pdb
• Descriptor: GDP-fucose protein O-fucosyltransferase 2,Spondin-1, 2-acetamido-2-deoxy-beta-D-glucopyranose, 1,2-ETHANEDIOL, ... (6 entities in total)
• Functional Keywords: pofut2, tsr, water molecules, protein o-fucosylation, transferase
• Biological source: Caenorhabditis elegans; More
• Total number of polymer chains: 2
• Total formula weight: 109010.33

Authors

Hurtado-Guerrero, R.,Merino, P. (deposition date: 2022-09-01, release date: 2022-10-26, Last modification date: 2024-10-09)

Primary citation

Sanz-Martinez, I.,Garcia-Garcia, A.,Tejero, T.,Hurtado-Guerrero, R.,Merino, P.
The Essential Role of Water Molecules in the Reaction Mechanism of Protein O-Fucosyltransferase 2.
Angew.Chem.Int.Ed.Engl., 61:e202213610-e202213610, 2022

PubMed Abstract: Protein O-fucosyltransferase 2 (PoFUT2) is an inverting glycosyltransferase (GT) that fucosylates thrombospondin repeats (TSRs) from group 1 and 2. PoFUT2 recognizes a large and diverse number of TSRs through a dynamic network of water-mediated interactions. By X-ray structural studies of C. elegans PoFUT2 complexed to a TSR of group 2, we demonstrate that this GT recognizes similarly the 3D structure of TSRs from both groups 1 and 2. Its active site is highly exposed to the solvent, suggesting that water molecules might also play an essential role in the fucosylation mechanism. We applied QM/MM methods using human PoFUT2 as a model, and found that HsPoFUT2 follows a classical S 2 reaction mechanism in which water molecules contribute to a great extent in facilitating the release of the leaving pyrophosphate unit, causing the H transfer from the acceptor nucleophile (Thr/Ser) to the catalytic base, which is the last event in the reaction. This demonstrates the importance of water molecules not only in recognition of the ligands but also in catalysis.

PubMed: 36260536
DOI: 10.1002/anie.202213610

Experimental method

X-RAY DIFFRACTION (2.13 Å)