| Structural and mechanistic studies of HpxO, a novel flavin adenine dinucleotide-dependent urate oxidase from Klebsiella pneumoniae. | |
| | |
MedLine Citation:
|
PMID: 23259842 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
HpxO is a flavin-dependent urate oxidase that catalyzes the hydroxylation of uric acid to 5-hydroxyisourate and functions in a novel pathway for purine catabolism found in Klebsiella pneumoniae. We have determined the structures of HpxO with and without uric acid at 2.0 and 2.2 Å, respectively. We have also determined the structure of the R204Q variant at 2.0 Å resolution in the absence of uric acid. The variant structure is very similar to that of wild-type HpxO except for the conformation of Arg103, which interacts with FAD in the variant but not in the wild-type structure. Interestingly, the R204Q variant results in the uncoupling of nicotinamide adenine dinucleotide oxidation from uric acid hydroxylation. This suggests that Arg204 facilitates the deprotonation of uric acid, activating it for the oxygen transfer. On the basis of these data, a mechanism for this reaction consisting of a nucleophilic attack of the urate anion on the flavin hydroperoxide resulting in the formation of 5-hydroxyisourate is proposed. |
| | |
Authors:
|
Katherine A Hicks; Seán E O'Leary; Tadhg P Begley; Steven E Ealick |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2013-01-09 |
Journal Detail:
|
Title: Biochemistry Volume: 52 ISSN: 1520-4995 ISO Abbreviation: Biochemistry Publication Date: 2013 Jan |
Date Detail:
|
Created Date: 2013-01-22 Completed Date: 2013-03-14 Revised Date: 2013-04-16 |
Medline Journal Info:
|
Nlm Unique ID: 0370623 Medline TA: Biochemistry Country: United States |
Other Details:
|
Languages: eng Pagination: 477-87 Citation Subset: IM |
Affiliation:
|
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA. |
| Data Bank Information | |
Bank Name/Acc. No.:
|
PDB/3RP6; 3RP7; 3RP8 |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Amino Acid Substitution Bacterial Proteins / chemistry*, genetics, metabolism* Biocatalysis Catalytic Domain Crystallography, X-Ray Flavin-Adenine Dinucleotide / chemistry, metabolism* Hydroxylation Kinetics Klebsiella pneumoniae / enzymology* Ligands Models, Molecular Molecular Conformation Mutagenesis, Site-Directed Mutant Proteins / chemistry, metabolism NAD / chemistry, metabolism Oxidation-Reduction Recombinant Proteins / chemistry, metabolism Urate Oxidase / chemistry*, genetics, metabolism* Uric Acid / analogs & derivatives, chemistry, metabolism |
| Grant Support | |
ID/Acronym/Agency:
|
GM073220/GM/NIGMS NIH HHS; P41 GM103403/GM/NIGMS NIH HHS; P41 RR015301/RR/NCRR NIH HHS; R01 GM073220/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/5-hydroxyisourate; 0/Bacterial Proteins; 0/Ligands; 0/Mutant Proteins; 0/Recombinant Proteins; 146-14-5/Flavin-Adenine Dinucleotide; 53-84-9/NAD; 69-93-2/Uric Acid; EC 1.7.3.3/Urate Oxidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Visualization and virtual screening of the chemical universe database GDB-17.
Next Document: Breastfeeding and its impact on daily life in women with type 1 diabetes during the first six months...