60708 20200221144154.0 doi 10.1038/nchembio.2019 sideral 94764 ART-2016-94764 eng (orcid)0000-0002-4680-937X Valero-González, J. A proactive role of water molecules in acceptor recognition by protein O-fucosyltransferase 2 2016 Access copy available to the general public Unrestricted 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. info:eu-repo/grantAgreement/ES/MINECO/CTQ2012-36365 info:eu-repo/grantAgreement/ES/MICINN/CTQ2013-44367-C2-2-P info:eu-repo/grantAgreement/ES/MICINN/BFU2010-19504 info:eu-repo/grantAgreement/EC/FP7/283570/EU/Transnational access and enhancement of integrated Biological Structure determination at synchrotron X-ray radiation facilities/BIOSTRUCT-X info:eu-repo/grantAgreement/ES/DGA/B89 info:eu-repo/semantics/openAccess All rights reserved http://www.europeana.eu/rights/rr-f/ 15.066 2016 BIOCHEMISTRY & MOLECULAR BIOLOGY 5 / 287 = 0.017 2016 Q1 T1 8.945 2016 Molecular Biology 2016 Q1 Cell Biology 2016 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion Leonhard-Melief, C. Lira-Navarrete, E. (orcid)0000-0003-0105-4337 Jiménez-Osés, G. Hernández-Ruiz, C. Pallarés, M.C. (orcid)0000-0003-3608-4720 Yruela, I. Universidad de Zaragoza Vasudevan, D. (orcid)0000-0001-7460-5916 Lostao, A. Universidad de Zaragoza Corzana, F. Takeuchi, H. Haltiwanger, R.S. (orcid)0000-0002-3122-9401 Hurtado-Guerrero, R. Universidad de Zaragoza 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 12, 4 (2016), 240-246 Nat. Chem. Biol. Nature Chemical Biology 1552-4450 2096935 http://zaguan.unizar.es/record/60708/files/texto_completo.pdf Postprint 48494 http://zaguan.unizar.es/record/60708/files/texto_completo.jpg?subformat=icon icon Postprint oai:zaguan.unizar.es:60708 articulos driver 2020-02-21-13:08:36 ARTICLE