Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/65116COMPARTIR / EXPORTAR:
 SHARE
 CORE
BASE
|
|
| Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
| Título: | Structural insights into the mechanism of protein O-fucosylation |
Autor: | Lira-Navarrete, Erandi; Tejero, Tomás CSIC ORCID; Merino, Pedro CSIC ORCID | Fecha de publicación: | 2011 | Editor: | Public Library of Science | Citación: | PLoS ONE 6(9): e25365 (2011) | Resumen: | Protein O-fucosylation is an essential post-translational modification, involved in the folding of target proteins and in the role of these target proteins during embryonic development and adult tissue homeostasis, among other things. Two different enzymes are responsible for this modification, Protein O-fucosyltransferase 1 and 2 (POFUT1 and POFUT2, respectively). Both proteins have been characterised biologically and enzymatically but nothing is known at the molecular or structural level. Here we describe the first crystal structure of a catalytically functional POFUT1 in an apo-form and in complex with GDP-fucose and GDP. The enzyme belongs to the GT-B family and is not dependent on manganese for activity. GDP-fucose/GDP is localised in a conserved cavity connected to a large solvent exposed pocket, which we show is the binding site of epidermal growth factor (EGF) repeats in the extracellular domain of the Notch Receptor. Through both mutational and kinetic studies we have identified which residues are involved in binding and catalysis and have determined that the Arg240 residue is a key catalytic residue. We also propose a novel S N1-like catalytic mechanism with formation of an intimate ion pair, in which the glycosidic bond is cleaved before the nucleophilic attack; and theoretical calculations at a DFT (B3LYP/6-31+G(d,p) support this mechanism. Thus, the crystal structure together with our mutagenesis studies explain the molecular mechanism of POFUT1 and provide a new starting point for the design of functional inhibitors to this critical enzyme in the future. | Descripción: | This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al. | Versión del editor: | http://dx.doi.org/10.1371/journal.pone.0025365 | URI: | http://hdl.handle.net/10261/65116 | DOI: | 10.1371/journal.pone.0025365 | Identificadores: | doi: 10.1371/journal.pone.0025365 issn: 1932-6203 |
| Aparece en las colecciones: | (ISQCH) Artículos |
Ficheros en este ítem:
| Fichero | Descripción | Tamaño | Formato | |
|---|---|---|---|---|
| Structural insights.pdf | 2,7 MB | Adobe PDF |  Visualizar/Abrir |
CORE Recommender
PubMed Central
Citations
48
checked on 02-may-2024
SCOPUSTM
Citations
82
checked on 14-may-2024
WEB OF SCIENCETM
Citations
77
checked on 26-feb-2024
Page view(s)
328
checked on 16-may-2024
Download(s)
244
checked on 16-may-2024
Google ScholarTM
Check
Altmetric
Altmetric
Artículos relacionados:
NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.