PUBLICATION
The identification of an integral membrane, cytochrome c urate oxidase completes the catalytic repertoire of a therapeutic enzyme
- Authors
- Doniselli, N., Monzeglio, E., Dal Palù, A., Merli, A., Percudani, R.
- ID
- ZDB-PUB-150909-1
- Date
- 2015
- Source
- Scientific Reports 5: 13798 (Journal)
- Registered Authors
- Keywords
- Data mining, Oxidoreductases
- MeSH Terms
-
- Amino Acid Sequence
- Gene Deletion
- Solubility
- Uric Acid/metabolism
- Cytochromes c/metabolism*
- Protein Interaction Domains and Motifs*
- Catalase/metabolism
- Phenotype
- Agrobacterium/genetics
- Agrobacterium/metabolism
- Ammonia/metabolism
- Gene Expression
- Membrane Proteins/chemistry
- Membrane Proteins/genetics
- Membrane Proteins/metabolism*
- Urate Oxidase/chemistry
- Urate Oxidase/genetics
- Urate Oxidase/metabolism*
- Oxidation-Reduction
- Molecular Sequence Data
- Sequence Alignment
- PubMed
- 26349049 Full text @ Sci. Rep.
Citation
Doniselli, N., Monzeglio, E., Dal Palù, A., Merli, A., Percudani, R. (2015) The identification of an integral membrane, cytochrome c urate oxidase completes the catalytic repertoire of a therapeutic enzyme. Scientific Reports. 5:13798.
Abstract
In living organisms, the conversion of urate into allantoin requires three consecutive enzymes. The pathway was lost in hominid, predisposing humans to hyperuricemia and gout. Among other species, the genomic distribution of the two last enzymes of the pathway is wider than that of urate oxidase (Uox), suggesting the presence of unknown genes encoding Uox. Here we combine gene network analysis with association rule learning to identify the missing urate oxidase. In contrast with the known soluble Uox, the identified gene (puuD) encodes a membrane protein with a C-terminal cytochrome c. The 8-helix transmembrane domain corresponds to DUF989, a family without similarity to known proteins. Gene deletion in a PuuD-encoding organism (Agrobacterium fabrum) abolished urate degradation capacity; the phenotype was fully restored by complementation with a cytosolic Uox from zebrafish. Consistent with H2O2 production by zfUox, urate oxidation in the complemented strain caused a four-fold increase of catalase. No increase was observed in the wild-type, suggesting that urate oxidation by PuuD proceeds through cytochrome c-mediated electron transfer. These findings identify a missing link in purine catabolism, assign a biochemical activity to a domain of unknown function (DUF989), and complete the catalytic repertoire of an enzyme useful for human therapy.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping