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dc.contributor.authorCarmona Rodríguez-Acosta, Fernandoes_ES
dc.contributor.authorMuñoz-Robles, Víctores_ES
dc.contributor.authorCuesta Martos, Rafaeles_ES
dc.contributor.authorGálvez Rodríguez, Natividad es_ES
dc.contributor.authorCapdevila, Mercèes_ES
dc.contributor.authorMaréchal, Jean-Didieres_ES
dc.contributor.authorDomínguez Vera, José Manuel es_ES
dc.date.accessioned2017-07-19T11:25:29Z
dc.date.available2017-07-19T11:25:29Z
dc.date.issued2014
dc.identifier.citationCarmona Rodríguez-Acosta, F.; et al. Monitoring lactoferrin iron levels by fluorescence resonance energy transfer: A combined chemical and computational study. Journal of Biological Inorganic Chemistry, 19(3): 439-447 (2014). [http://hdl.handle.net/10481/47218]es_ES
dc.identifier.issn0949-8257
dc.identifier.issn1432-1327
dc.identifier.urihttp://hdl.handle.net/10481/47218
dc.description.abstractThree forms of lactoferrin (Lf) that differed in their levels of iron loading (Lf, LfFe, and LfFe2) were simultaneously labeled with the fluorophores AF350 and AF430. All three resulting fluorescent lactoferrins exhibited fluorescence resonance energy transfer (FRET), but they all presented different FRET patterns. Whereas only partial FRET was observed for Lf and LfFe, practically complete FRET was seen for the holo form (LfFe2). For each form of metal-loaded lactoferrin, the AF350–AF430 distance varied depending on the protein conformation, which in turn depended on the level of iron loading. Thus, the FRET patterns of these lactoferrins were found to correlate with their iron loading levels. In order to gain greater insight into the number of fluorophores and the different FRET patterns observed (i.e., their iron levels), a computational analysis was performed. The results highlighted a number of lysines that have the greatest influence on the FRET profile. Moreover, despite the lack of an X-ray structure for any LfFe species, our study also showed that this species presents modified subdomain organization of the N-lobe, which narrows its iron-binding site. Complete domain rearrangement occurs during the LfFe to LfFe2 transition. Finally, as an example of the possible applications of the results of this study, we made use of the FRET fingerprints of these fluorescent lactoferrins to monitor the interaction of lactoferrin with a healthy bacterium, namely Bifidobacterium breve. This latter study demonstrated that lactoferrin supplies iron to this bacterium, and suggested that this process occurs with no protein internalization.en_EN
dc.description.sponsorshipThis work was supported by MINECO and FEDER (projects CTQ2012-32236, CTQ2011-23336, and BIO2012-39682-C02-02) and BIOSEARCH SA. F.C. and V.M.R. are grateful to the Spanish MINECO for FPI fellowships.es_ES
dc.language.isoenges_ES
dc.publisherSpringer Verlanges_ES
dc.subjectFRETen_EN
dc.subjectLactoferrinen_EN
dc.subjectDockingen_EN
dc.subjectIron metabolismen_EN
dc.subjectProtein-ligand dockingen_EN
dc.subjectStructural analysisen_EN
dc.titleMonitoring lactoferrin iron levels by fluorescence resonance energy transfer: A combined chemical and computational studyen_EN
dc.typeinfo:eu-repo/semantics/articleen_EN
dc.rights.accessRightsinfo:eu-repo/semantics/openAccessen_EN
dc.identifier.doi10.1007/s00775-013-1088-z


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