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dc.contributor.authorLimones Herrero, Daniel
dc.contributor.authorPérez Ruiz, Raúl
dc.contributor.authorLence Quintana, Emilio José
dc.contributor.authorGonzález Bello, Concepción
dc.contributor.authorMiranda, Miguel A.
dc.contributor.authorJiménez, Consuelo M.
dc.date.accessioned2018-07-02T11:42:41Z
dc.date.available2018-07-02T11:42:41Z
dc.date.issued2017-01-05
dc.identifier.citationLimones-Herrero, D., Pérez-Ruiz, R., Lence, E., González-Bello, C., Miranda, M., & Jiménez, M. (2017). Mapping a protein recognition centre with chiral photoactive ligands. An integrated approach combining photophysics, reactivity, proteomics and molecular dynamics simulation studies. Chemical Science, 8(4), 2621-2628. doi: 10.1039/c6sc04900a
dc.identifier.urihttp://hdl.handle.net/10347/16922
dc.description.abstractA multidisciplinary strategy to obtain structural information on the intraprotein region is described here. As probe ligands, (S)- and (R)-CPFMe (the methyl esters of the chiral drug carprofen) have been selected, while bovine α1-acid glycoprotein (BAAG) has been chosen as a biological host. The procedure involves the separate irradiation of the BAAG/(S)-CPFMe and BAAG/(R)-CPFMe complexes, coupled with fluorescence spectroscopy, laser flash photolysis, proteomic analysis, docking and molecular dynamics simulations. Thus, irradiation of the BAAG/CPFMe complexes at λ = 320 nm was followed by fluorescence spectroscopy. The intensity of the emission band obtained after irradiation indicated photodehalogenation, whereas its structureless shape suggested covalent binding of the resulting radical CBZMe˙ to the biopolymer. After gel filtration chromatography, the spectra still displayed emission, in agreement with covalent attachment of CBZMe˙ to BAAG. Stereodifferentiation was observed in this process. After trypsin digestion and ESI-MS/MS, the incorporation of CBZMe was detected at Phe68. Docking and molecular dynamics simulation studies, which were carried out using a homology model of BAAG, reveal that the closer proximity of the aromatic moiety of the (S)-enantiomer to the phenyl group of Phe68 would be responsible for the experimentally observed, more effective chemical modification of the protein. The proposed tridimensional structure of BAAG covalently modified by the two enantiomers is also provided. In principle, this approach can be extended to a variety of protein/ligand complexes
dc.description.sponsorshipFinancial support from the Spanish Ministry of Economy and Competitiveness (CTQ2013-47872-C2-1-P, SAF2013-42899-R, BES-2011-043706), Generalitat Valenciana (PROMETEOII/2013/005), Instituto de Salud Carlos III (RD12/0013/0009), Xunta de Galicia (GRC2013-041), the Consellería de Cultura, Educación e Ordenación Universitaria (Centro singular de investigación de Galicia accreditation 2016-2019, ED431G/09) and the European Regional Development Fund (ERDF) is gratefully acknowledged. E. L. thanks the Xunta de Galicia for a postdoctoral fellowship. We are grateful to the Centro de Supercomputación de Galicia (CESGA) for use of the Finis Terrae II supercomputer. The proteomic analysis was performed in the proteomics facility of SCSIE University of Valencia that belongs to ProteoRed PRB2-ISCIII and is supported by grant PT13/0001, of the PE I+D+i 2013–2016, funded by ISCIII and FEDER
dc.language.isoeng
dc.publisherRoyal Society of Chemistry
dc.relationinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2013-42899-R/ES/DESARROLLO DE NUEVOS ANTIBIOTICOS PARA EL TRATAMIENTO DE INFECCIONES BACTERIANAS RESISTENTES: METABOLISMO, RESISTENCIA Y COMUNICACION CELULA-CELULA
dc.relationinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2013-47872-C2-1-P/ES/METABOLITOS FOTOACTIVOS
dc.rightsThis article is licensed under a Creative Commons Attribution 3.0 Unported Licence
dc.rights.urihttps://creativecommons.org/licenses/by/3.0/
dc.titleMapping a protein recognition centre with chiral photoactive ligands. An integrated approach combining photophysics, reactivity, proteomics and molecular dynamics simulation studies
dc.typeinfo:eu-repo/semantics/article
dc.identifier.DOI10.1039/C6SC04900A
dc.relation.publisherversionhttps://doi.org/10.1039/C6SC04900A
dc.type.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.e-issn2041-6539
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.contributor.affiliationUniversidade de Santiago de Compostela. Centro de Investigación en Química Biolóxica e Materiais Moleculares
dc.contributor.affiliationUniversidade de Santiago de Compostela. Departamento de Química Orgánica
dc.description.peerreviewedSI


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