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dc.contributor.authorMiki, Yuta-
dc.contributor.authorCalviño, Fabiola R.-
dc.contributor.authorPogni, Rebecca-
dc.contributor.authorGiansanti, Stefania-
dc.contributor.authorRuiz-Dueñas, F. J.-
dc.contributor.authorMartínez, María Jesús-
dc.contributor.authorBasosi, Riccardo-
dc.contributor.authorRomero, Antonio-
dc.contributor.authorMartínez, Ángel T.-
dc.date.issued2011-04-29-
dc.identifier.citationJournal of Biological Chemistry 286: 5525-15534(2011)es_ES
dc.identifier.issn0021-9258-
dc.identifier.uri10261/49320-
dc.description12 p.-8 fig.-3 tab.-1 fig. supl.es_ES
dc.description.abstractTrametes cervina lignin peroxidase (LiP) is a unique enzyme lacking the catalytic tryptophan strictly conserved in all other LiPs and versatile peroxidases (more than 30 sequences available). Recombinant T. cervina LiP and site-directed variants were investigated by crystallographic, kinetic, and spectroscopic techniques. The crystal structure shows three substrate oxidation site candidates involving His-170, Asp-146, and Tyr-181. Steady-state kinetics for oxidation of veratryl alcohol (the typical LiP substrate) by variants at the above three residues reveals a crucial role of Tyr-181 in LiP activity. Moreover, assays with ferrocytochrome c show that its ability to oxidize large molecules (a requisite property for oxidation of the lignin polymer) originates in Tyr-181. This residue is also involved in the oxidation of 1,4-dimethoxybenzene, a reaction initiated by the one-electron abstraction with formation of substrate cation radical, as described for the well known Phanerochaete chrysosporium LiP. Detailed spectroscopic and kinetic investigations, including low temperature EPR, show that the porphyrin radical in the two-electron activated T. cervina LiP is unstable and rapidly receives one electron from Tyr-181, forming a catalytic protein radical, which is identified as an H-bonded neutral tyrosyl radical. The crystal structure reveals a partially exposed location of Tyr-181, compatible with its catalytic role, and several neighbor residues probably contributing to catalysis: (i) by enabling substrate recognition by aromatic interactions; (ii) by acting as proton acceptor/donor from Tyr-181 or H-bonding the radical form; and (iii) by providing the acidic environment that would facilitate oxidation. This is the first structure-function study of the only ligninolytic peroxidase described to date that has a catalytic tyrosinees_ES
dc.description.sponsorshipThis work was supported by the Spanish projects BIO2008-01533 (RAPERO) and BFU2008-02595, the European projects BIORENEW (NMP2-CT-2006-026456) and PEROXICATS (KBBE-2010-4-265397), and the Italian Ministry of University and Research (MIUR) project 20072R7WWAes_ES
dc.language.isoenges_ES
dc.publisherAmerican Society for Biochemistry and Molecular Biologyes_ES
dc.relationBIORENEW (NMP2-CT-2006-026456) and PEROXICATS (KBBE-2010-4-265397), and the Italian Ministry of University and Research (MIUR) project 20072R7WWAes_ES
dc.rightsopenAccesses_ES
dc.subjectCrystal Structurees_ES
dc.subjectElectron Paramagnetic Resonance (EPR)es_ES
dc.subjectElectron Transferes_ES
dc.subjectEnzyme Kineticses_ES
dc.subjectEnzyme Mechanismses_ES
dc.subjectSite-directed Mutagenesises_ES
dc.subjectTrametes cervinaes_ES
dc.subjectCompounds I and IIes_ES
dc.subjectLignin Peroxidasees_ES
dc.subjectTyrosyl Radicales_ES
dc.titleCrystallographic, kinetic, and spectroscopic study of the first ligninolytic peroxidase presenting a catalytic tyrosinees_ES
dc.typeartículoes_ES
dc.identifier.doi10.1074/jbc.M111.220996-
dc.description.peerreviewedPeer reviewedes_ES
dc.relation.publisherversionhttp://dx.doi.org/10.1074/jbc.M111.220996es_ES
dc.identifier.e-issn1083-351X286-
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