Site-directed mutations and kinetic studies show key residues involved in alkylammonium interactions and reveal two sites for phosphorylcholine in Pseudomonas aeruginosa phosphorylcholine phosphatase

dc.creatorBeassoni, P.R.
dc.creatorOtero, L.H.
dc.creatorBoetsch, C.
dc.creatorDomenech, C.E.
dc.creatorGonzalez-Nilo, F.D.
dc.creatorLisa, A.T.
dc.date2012-09-06T21:03:10Z
dc.date2012-09-06T21:03:10Z
dc.date2011-07
dc.date.accessioned2017-03-07T14:58:18Z
dc.date.available2017-03-07T14:58:18Z
dc.identifierBIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS Volume: 1814 Issue: 7 Pages: 858-863 DOI: 10.1016/j.bbapap.2011.04.003
dc.identifier1570-9639
dc.identifierhttp://dspace.utalca.cl/handle/1950/8812
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/375747
dc.descriptionGonzalez-Nilo, FD (Gonzalez-Nilo, Fernado D.)Univ Talca, Ctr Bioinformat & Simulac Mol, Talca, Chile
dc.descriptionPseudomonas aeruginosa phosphorylcholine phosphatase (PchP) catalyzes the hydrolysis of phosphorylcholine (Pcho) to produce choline and inorganic phosphate. PchP belongs to the haloacid dehalogenase superfamily (HAD) and possesses the three characteristic motifs of this family: motif I ((31)D and (33)D), motif II ((166)S), and motif III ((242)K, (261)G, (262)D and (267)D), which fold to form the catalytic site that binds the metal ion and the phosphate moiety of Pcho. Based on comparisons to the PHOSPHO1 and PHOSPHO2 human enzymes and the choline-binding proteins of Gram-(+) bacteria, we selected residues (42)E and (43)E and the aromatic triplet (82)YYY(84) for site-directed mutagenesis to study the interactions with Pcho and p-nitrophenylphosphate as substrates of PchP. Because mutations in (42)E, (43)E and the three tyrosine residues affect both the substrate affinity and the inhibitory effect produced by high Pcho concentrations, we postulate that two sites, one catalytic and one inhibitory, are present in PchP and that they are adjacent and share residues. (C) 2011 Elsevier B.V. All rights reserved.
dc.languageen
dc.publisherELSEVIER SCIENCE BV, PO BOX 211
dc.subjectPseudomonas aeruginosa
dc.subjectPhosphorylcholine
dc.subjectCholine
dc.subjectSite-directed mutagenesis
dc.subjectHAD superfamily
dc.subjectSubstrate inhibition
dc.titleSite-directed mutations and kinetic studies show key residues involved in alkylammonium interactions and reveal two sites for phosphorylcholine in Pseudomonas aeruginosa phosphorylcholine phosphatase
dc.typeArtículos de revistas


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