A proactive role of water molecules in acceptor recognition by protein O-fucosyltransferase 2

Valero-González, J. ; Leonhard-Melief, C. ; Lira-Navarrete, E. ; Jiménez-Osés, G. ; Hernández-Ruiz, C. ; Pallarés, M.C. ; Yruela, I. (Universidad de Zaragoza) ; Vasudevan, D. ; Lostao, A. (Universidad de Zaragoza) ; Corzana, F. ; Takeuchi, H. ; Haltiwanger, R.S. ; Hurtado-Guerrero, R. (Universidad de Zaragoza)

Resumen: 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.

Idioma: Inglés

DOI: 10.1038/nchembio.2019

Año: 2016

Publicado en: Nature Chemical Biology 12, 4 (2016), 240-246
ISSN: 1552-4450

Factor impacto JCR: 15.066 (2016)

Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 5 / 287 = 0.017 (2016) - Q1 - T1

Factor impacto SCIMAGO: 8.945 - Molecular Biology (Q1) - Cell Biology (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/B89
Financiación: 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
Financiación: info:eu-repo/grantAgreement/ES/MICINN/BFU2010-19504
Financiación: info:eu-repo/grantAgreement/ES/MICINN/CTQ2013-44367-C2-2-P
Financiación: info:eu-repo/grantAgreement/ES/MINECO/CTQ2012-36365

Tipo y forma: Artículo (PostPrint)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)