| http://purl.uniprot.org/citations/15226302 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/15226302 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/15226302 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Citation |
| http://purl.uniprot.org/citations/15226302 | http://www.w3.org/2000/01/rdf-schema#comment | "WbpA (PA3159) is an enzyme involved in the biosynthesis of unusual di-N-acetyl-d-mannosaminuronic acid-derived sugar nucleotides found in the O antigen of Pseudomonas aeruginosa PAO1 (serotype O5). The wbpA gene that encodes this enzyme was cloned into pET-28a, overexpressed as a histidine-tagged fusion protein, and purified by nickel chelation chromatography. Capillary electrophoresis was used to examine substrate conversion by WbpA, and the data revealed that WbpA is a UDP-N-acetyl-D-glucosamine 6-dehydrogenase (EC 1.1.1.136), which uses NAD(+) as a coenzyme. The enzyme reaction product was purified by HPLC and analyzed using NMR spectroscopy. Our results showed unequivocally that the product of the WbpA reaction is UDP-N-acetyl-d-glucosaminuronic acid. WbpA requires either NH(4)(+) or K(+) for activity and the accompanying anions exert secondary effects on activity consistent with their ranking in the Hofmeister series. Kinetic analysis showed positive cooperativity with respect to UDP-N-acetyl-d-glucosamine binding with a K(0.5) of 94 microM, a k(cat) of 86 min(-1), and a Hill coefficient of 1.8. In addition, WbpA has a K(0.5) for NAD(+) of 220 microM, a k(cat) of 86 min(-1), and a Hill coefficient of 1.1. The oligomerization state of WbpA was analyzed by gel filtration, dynamic light scattering, and analytical ultracentrifugation, with all three techniques indicating that WbpA exists as a trimer in solution. However, tertiary structure predictions suggested a tetramer, which was supported by data from transmission electron microscopy. The electron micrograph of negatively stained WbpA samples revealed structures with 4-fold symmetry."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.org/dc/terms/identifier | "doi:10.1074/jbc.m404749200"xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.org/dc/terms/identifier | "doi:10.1074/jbc.m404749200"xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Brisson J.R."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Brisson J.R."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Daniels C."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Daniels C."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Wenzel C.Q."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Wenzel C.Q."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Lam J.S."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Lam J.S."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Miller W.L."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Miller W.L."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Larocque S."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/author | "Larocque S."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/date | "2004"xsd:gYear |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/date | "2004"xsd:gYear |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/name | "J. Biol. Chem."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/name | "J. Biol. Chem."xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/pages | "37551-37558"xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/pages | "37551-37558"xsd:string |
| http://purl.uniprot.org/citations/15226302 | http://purl.uniprot.org/core/title | "Biochemical characterization of WbpA, a UDP-N-acetyl-D-glucosamine 6-dehydrogenase involved in O-antigen biosynthesis in Pseudomonas aeruginosa PAO1."xsd:string |