Journal
NATURE CHEMICAL BIOLOGY
Volume 12, Issue 4, Pages 240-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.2019
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Funding
- Agencia Aragonesa para la Investigacion y Desarrollo (ARAID)
- Ministerio de Economia y Competitividad (MEC) [BFU2010-19504, CTQ2013-44367-C2-2-P, CTQ2012-36365]
- US National Institutes of Health [GM061126, CA123071]
- Ramon y Cajal Programme
- Diputacion General de Aragon (DGA) [B89]
- EU Seventh Framework Programme under BioStruct-X [283570, BIOSTRUCTX 5186]
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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.
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