5FOE

Crystal structure of the C. elegans Protein O-fucosyltransferase 2 (CePOFUT2) double mutant (R298K-R299K) in complex with GDP and the human TSR1 from thrombospondin 1

Summary for 5FOE

• Entry DOI: 10.2210/pdb5foe/pdb
• Descriptor: GDP-fucose protein O-fucosyltransferase 2,Thrombospondin-1, 2-acetamido-2-deoxy-beta-D-glucopyranose, GUANOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• Functional Keywords: transferase, pofut2, waters, fusion protein, afm, itc, glycosyltransferase, site-directed mutagenesis, molecular dynamics, tsr1
• Biological source: Caenorhabditis elegans; More
• Total number of polymer chains: 2
• Total formula weight: 116979.95

Authors

Valero-Gonzalez, J.,Leonhard-Melief, C.,Lira-Navarrete, E.,Jimenez-Oses, G.,Hernandez-Ruiz, C.,Pallares, M.C.,Yruela, I.,Vasudevan, D.,Lostao, A.,Corzana, F.,Takeuchi, H.,Haltiwanger, R.S.,Hurtado-Guerrero, R. (deposition date: 2015-11-19, release date: 2016-01-27, Last modification date: 2024-11-20)

Primary citation

Valero-Gonzalez, J.,Leonhard-Melief, C.,Lira-Navarrete, E.,Jimenez-Oses, G.,Hernandez-Ruiz, C.,Pallares, M.C.,Yruela, I.,Vasudevan, D.,Lostao, A.,Corzana, F.,Takeuchi, H.,Haltiwanger, R.S.,Hurtado-Guerrero, R.
A Proactive Role of Water Molecules in Acceptor Recognition of Thrombospondin Type 1 Repeats by Protein-O-Fucosyltransferase 2
Nat.Chem.Biol., 12:240-, 2016

PubMed 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.

PubMed: 26854667
DOI: 10.1038/NCHEMBIO.2019

Experimental method

X-RAY DIFFRACTION (1.98 Å)