2w23

STRUCTURE OF MUTANT W169Y OF PLEUROTUS ERYNGII VERSATILE PEROXIDASE (VP)

spinqualitylabelsall models 2w23, resolution 1.94Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 2w23 is a 1 chain structure with sequence from Pleurotus eryngii. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: CA, GOL, HEM, SO4 Related: 2boq, 2vka Activity: Versatile peroxidase, with EC number 1.11.1.16 Resources: FirstGlance, OCA, RCSB, PDBsum Function [VPL2_PLEER] A versatile ligninolytic peroxidase that combines the substrate specificity characteristics of the two other ligninolytic peroxidases, manganese peroxidase and lignin peroxidase. Evolutionary Conservation Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf. Publication Abstract from PubMed Lignin-degrading peroxidases, a group of biotechnologically-interesting enzymes,?oxidize high redox-potential aromatics via an exposed protein radical. Low-temperature EPR of Pleurotus eryngii versatile peroxidase (VP) revealed, for the first time in a fungal peroxidase, the presence of a tryptophanyl radical in both the two-electron (VPI) and one-electron (VPII) activated forms of the enzyme. Site-directed mutagenesis was used to substitute this tryptophan (Trp164) by tyrosine and histidine residues. No changes in the crystal structure were observed indicating that the modified behavior was due exclusively to the mutations introduced. EPR revealed the formation of tyrosyl radicals in both VPI and VPII of the W164Y variant. However, no protein radical was detected in the W164H variant, whose VPI spectrum indicated a porphyrin radical identical to that of the inactive W164S. Stopped-flow spectrophotometry showed that the W164Y mutation reduced 10-fold the apparent second-order rate constant for VPI reduction (k2app) by veratryl alcohol (VA), compared with over 50-fold reduction in W164S, revealing some catalytic activity of the tyrosine radical. Its first-order rate constant (k2) was more affected than the dissociation constant (KD2). Moreover, VPII reduction by VA was impaired by the above mutations revealing that the Trp164 radical was involved in catalysis by both VPI and VPII. The low first-order rate constant (k3) values were similar for the W164Y, W164H and W164S variants, indicating that the tyrosyl radical in VPII was not able to oxidize VA (in contrast with that observed for VPI). VPII self-reduction was also suppressed revealing that Trp164 is involved in this autocatalytic process. Protein radicals in fungal versatile peroxidase: Catalytic tryptophan radical in both Compound-I and Compound-II and studies on W164Y, W164H and W164S variants.,Ruiz-Duenas FJ, Pogni R, Morales M, Giansanti S, Mate MJ, Romero A, Martinez MJ, Basosi R, Martinez AT J Biol Chem. 2009 Jan 21. PMID:19158088 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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