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dc.contributor.authorGarcia-Guerrero, Maria C.es_ES
dc.contributor.authorGarcía-Pardo, Javieres_ES
dc.contributor.authorBerenguer, Estheres_ES
dc.contributor.authorFernandez-Alvarez, Robertoes_ES
dc.contributor.authorBarfi, Gifty B.es_ES
dc.contributor.authorLyons, Peter J.es_ES
dc.contributor.authorAvilés, Francesc Xavieres_ES
dc.contributor.authorHuber, Robertes_ES
dc.contributor.authorLorenzo, Juliaes_ES
dc.contributor.authorReverter, Davides_ES
dc.date.accessioned2020-01-29T09:53:26Z-
dc.date.available2020-01-29T09:53:26Z-
dc.date.issued2018-
dc.identifier.citationProceedings of the National Academy of Sciences 115(17): E3932-E3939 (2018)es_ES
dc.identifier.issn0027-8424-
dc.identifier.urihttp://hdl.handle.net/10261/199151-
dc.description.abstractHuman metallocarboxypeptidase O (hCPO) is a recently discovered digestive enzyme localized to the apical membrane of intestinal epithelial cells. Unlike pancreatic metallocarboxypeptidases, hCPO is glycosylated and produced as an active enzyme with distinctive substrate specificity toward C-terminal (C-t) acidic residues. Here we present the crystal structure of hCPO at 1.85-Å resolution, both alone and in complex with a carboxypeptidase inhibitor (NvCI) from the marine snail Nerita versicolor. The structure provides detailed information regarding determinants of enzyme specificity, in particular Arg275, placed at the bottom of the substrate-binding pocket. This residue, located at “canonical” position 255, where it is Ile in human pancreatic carboxypeptidases A1 (hCPA1) and A2 (hCPA2) and Asp in B (hCPB), plays a dominant role in determining the preference of hCPO for acidic C-t residues. Site-directed mutagenesis to Asp and Ala changes the specificity to C-t basic and hydrophobic residues, respectively. The single-site mutants thus faithfully mimic the enzymatic properties of CPB and CPA, respectively. hCPO also shows a preference for Glu over Asp, probably as a consequence of a tighter fitting of the Glu side chain in its S1′ substrate-binding pocket. This unique preference of hCPO, together with hCPA1, hCPA2, and hCPB, completes the array of C-t cleavages enabling the digestion of the dietary proteins within the intestine. Finally, in addition to activity toward small synthetic substrates and peptides, hCPO can also trim C-t extensions of proteins, such as epidermal growth factor, suggesting a role in the maturation and degradation of growth factors and bioactive peptides.es_ES
dc.description.sponsorshipThis work was funded by the Spanish Ministry of Innovation and Competitiveness Grants BFU2015-66417-P Ministry of Economy and Competitiveness (MINECO) and the European Fund for Regional Development (FEDER) (to D.R.) and BIO2016-78057-R (to F.X.A.); and by a Faculty Research grant from Andrews University (to P.J.L.). J.G.-P. and M.C.G.-G. were supported by PhD Fellowships BES-2011-044872 and FPU12/06137, respectively, from MINECO. J.G.-P. was supported by short-term Fellowship ASTF 603–2015 from the European Molecular Biology Organization.es_ES
dc.language.isoenges_ES
dc.publisherNational Academy of Sciences (U.S.)es_ES
dc.relationMINECO/ICTI2013-2016/BFU2015-66417-Pes_ES
dc.relationMINECO/ICTI2013-2016/BIO2016-78057-Res_ES
dc.relation.isversionofPublisher's versiones_ES
dc.rightsopenAccesses_ES
dc.subjectCarboxypeptidasees_ES
dc.subjectProtein digestiones_ES
dc.subjectCrystal structurees_ES
dc.subjectAcidic proteasees_ES
dc.titleCrystal structure and mechanism of human carboxypeptidase O: Insights into its specific activity for acidic residueses_ES
dc.typeartículoes_ES
dc.identifier.doi10.1073/pnas.1803685115-
dc.description.peerreviewedPeer reviewedes_ES
dc.relation.publisherversionhttps://doi.org/10.1073/pnas.1803685115es_ES
dc.identifier.e-issn1091-6490-
dc.contributor.funderMinisterio de Economía y Competitividad (España)es_ES
dc.contributor.funderEuropean Commissiones_ES
dc.contributor.funderUniversity of St Andrewses_ES
dc.contributor.funderEuropean Molecular Biology Laboratoryes_ES
dc.relation.csices_ES
oprm.item.hasRevisionno ko 0 false*
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000780es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000740es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/100013060es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000780es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000740es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100003329es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/100013060es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100003329es_ES
dc.contributor.orcidGarcía-Pardo, Javier [0000-0001-9179-6371]es_ES
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