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Protein

FAD-dependent urate hydroxylase

Gene

hpxO

Organism
Klebsiella pneumoniae subsp. pneumoniae (strain ATCC 700721 / MGH 78578)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the hydroxylation of urate to 5-hydroxyisourate (HIU).1 Publication

Catalytic activityi

Urate + NADH + O2 = 5-hydroxyisourate + NAD+ + H2O.1 Publication

Cofactori

FAD1 Publication

Kineticsi

kcat is 42 sec(-1).1 Publication

  1. KM=42 µM for urate1 Publication

    Pathwayi: urate degradation

    This protein is involved in the pathway urate degradation, which is part of Purine metabolism.1 Publication
    View all proteins of this organism that are known to be involved in the pathway urate degradation and in Purine metabolism.

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei11 – 111FAD; via amide nitrogenCombined sources1 Publication
    Binding sitei43 – 431FADCombined sources1 Publication
    Binding sitei125 – 1251FAD; via amide nitrogen and carbonyl oxygenCombined sources1 Publication
    Binding sitei178 – 1781SubstrateCombined sources1 Publication
    Binding sitei204 – 2041SubstrateCombined sources1 Publication
    Sitei204 – 2041Involved in substrate activation for the transfer of oxygen from the flavin hydroperoxide1 Publication
    Binding sitei285 – 2851FADCombined sources1 Publication

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi30 – 312FADCombined sources1 Publication
    Nucleotide bindingi295 – 2995FADCombined sources1 Publication

    GO - Molecular functioni

    • FAD binding Source: UniProtKB
    • oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen Source: UniProtKB
    • urate oxidase activity Source: CACAO

    GO - Biological processi

    • purine nucleobase metabolic process Source: UniProtKB-KW
    • urate catabolic process Source: UniProtKB
    Complete GO annotation...

    Keywords - Molecular functioni

    Monooxygenase, Oxidoreductase

    Keywords - Biological processi

    Purine metabolism

    Keywords - Ligandi

    FAD, Flavoprotein, NAD

    Enzyme and pathway databases

    BioCyciKPNE272620:GKDC-1663-MONOMER.
    BRENDAi1.14.13.113. 2814.
    UniPathwayiUPA00394.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    FAD-dependent urate hydroxylase1 Publication (EC:1.14.13.1131 Publication)
    Alternative name(s):
    FAD-dependent urate oxidase1 Publication
    Gene namesi
    Name:hpxO1 Publication
    Ordered Locus Names:KPN78578_16330
    ORF Names:KPN_01663
    OrganismiKlebsiella pneumoniae subsp. pneumoniae (strain ATCC 700721 / MGH 78578)
    Taxonomic identifieri272620 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeKlebsiella
    Proteomesi
    • UP000000265 Componenti: Chromosome

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi204 – 2041R → Q: 160-fold decrease in catalytic activity. Results in the uncoupling of the NADH oxidation and urate hydroxylation reactions. 1 Publication
    Mutagenesisi216 – 2161Y → F: 5-fold decrease in catalytic activity. 2-fold decrease in affinity for urate and increase in affinity for NADH. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 384384FAD-dependent urate hydroxylasePRO_0000418845Add
    BLAST

    Interactioni

    Subunit structurei

    Monomer.1 Publication

    Protein-protein interaction databases

    STRINGi272620.KPN_01663.

    Structurei

    Secondary structure

    1
    384
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi2 – 65Combined sources
    Helixi10 – 2112Combined sources
    Beta strandi25 – 339Combined sources
    Beta strandi41 – 444Combined sources
    Helixi46 – 549Combined sources
    Helixi58 – 647Combined sources
    Beta strandi70 – 756Combined sources
    Turni76 – 783Combined sources
    Beta strandi81 – 866Combined sources
    Helixi88 – 947Combined sources
    Beta strandi99 – 1024Combined sources
    Helixi103 – 11412Combined sources
    Helixi116 – 1183Combined sources
    Beta strandi119 – 1224Combined sources
    Beta strandi125 – 1317Combined sources
    Beta strandi134 – 1396Combined sources
    Beta strandi144 – 1529Combined sources
    Helixi161 – 1655Combined sources
    Beta strandi172 – 18413Combined sources
    Turni187 – 1893Combined sources
    Beta strandi194 – 2007Combined sources
    Beta strandi203 – 2108Combined sources
    Turni211 – 2133Combined sources
    Beta strandi214 – 2229Combined sources
    Turni231 – 2333Combined sources
    Helixi234 – 2418Combined sources
    Turni242 – 2443Combined sources
    Helixi247 – 2559Combined sources
    Helixi258 – 2603Combined sources
    Beta strandi262 – 2687Combined sources
    Beta strandi280 – 2823Combined sources
    Helixi284 – 2863Combined sources
    Helixi292 – 2943Combined sources
    Helixi297 – 31317Combined sources
    Helixi318 – 34730Combined sources
    Turni348 – 3514Combined sources
    Helixi352 – 36312Combined sources
    Helixi368 – 37912Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3RP6X-ray2.20A1-384[»]
    3RP7X-ray2.04A1-384[»]
    3RP8X-ray1.97A1-384[»]
    ProteinModelPortaliA6T923.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni216 – 2183Substrate bindingCombined sources1 Publication

    Sequence similaritiesi

    Phylogenomic databases

    eggNOGiENOG41061H0. Bacteria.
    COG0654. LUCA.
    HOGENOMiHOG000166536.
    KOiK16839.
    OMAiRIINGLC.
    OrthoDBiEOG6TR0FF.

    Family and domain databases

    Gene3Di3.50.50.60. 2 hits.
    InterProiIPR002938. FAD-bd.
    IPR023753. FAD/NAD-binding_dom.
    [Graphical view]
    PfamiPF01494. FAD_binding_3. 1 hit.
    [Graphical view]
    SUPFAMiSSF51905. SSF51905. 2 hits.

    Sequencei

    Sequence statusi: Complete.

    A6T923-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MKAIVIGAGI GGLSAAVALK QSGIDCDVYE AVKEIKPVGA AISVWPNGVK
    60 70 80 90 100
    CMAHLGMGDI METFGGPLRR MAYRDFRSGE NMTQFSLAPL IERTGSRPCP
    110 120 130 140 150
    VSRAELQREM LDYWGRDSVQ FGKRVTRCEE DADGVTVWFT DGSSASGDLL
    160 170 180 190 200
    IAADGSHSAL RPWVLGFTPQ RRYAGYVNWN GLVEIDEALA PGDQWTTFVG
    210 220 230 240 250
    EGKRVSLMPV SAGRFYFFFD VPLPAGLAED RDTLRADLSR YFAGWAPPVQ
    260 270 280 290 300
    KLIAALDPQT TNRIEIHDIE PFSRLVRGRV ALLGDAGHST TPDIGQGGCA
    310 320 330 340 350
    AMEDAVVLGA VFRQTRDIAA ALREYEAQRC DRVRDLVLKA RKRCDITHGK
    360 370 380
    DMQLTEAWYQ ELREETGERI INGMCDTILS GPLG
    Length:384
    Mass (Da):42,107
    Last modified:August 21, 2007 - v1
    Checksum:i4B656520269C3535
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    CP000647 Genomic DNA. Translation: ABR77094.1.
    RefSeqiWP_002904783.1. NC_009648.1.

    Genome annotation databases

    EnsemblBacteriaiABR77094; ABR77094; KPN_01663.
    KEGGikpn:KPN_01663.
    PATRICi20457612. VBIKlePne13394_1694.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    CP000647 Genomic DNA. Translation: ABR77094.1.
    RefSeqiWP_002904783.1. NC_009648.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3RP6X-ray2.20A1-384[»]
    3RP7X-ray2.04A1-384[»]
    3RP8X-ray1.97A1-384[»]
    ProteinModelPortaliA6T923.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi272620.KPN_01663.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiABR77094; ABR77094; KPN_01663.
    KEGGikpn:KPN_01663.
    PATRICi20457612. VBIKlePne13394_1694.

    Phylogenomic databases

    eggNOGiENOG41061H0. Bacteria.
    COG0654. LUCA.
    HOGENOMiHOG000166536.
    KOiK16839.
    OMAiRIINGLC.
    OrthoDBiEOG6TR0FF.

    Enzyme and pathway databases

    UniPathwayiUPA00394.
    BioCyciKPNE272620:GKDC-1663-MONOMER.
    BRENDAi1.14.13.113. 2814.

    Family and domain databases

    Gene3Di3.50.50.60. 2 hits.
    InterProiIPR002938. FAD-bd.
    IPR023753. FAD/NAD-binding_dom.
    [Graphical view]
    PfamiPF01494. FAD_binding_3. 1 hit.
    [Graphical view]
    SUPFAMiSSF51905. SSF51905. 2 hits.
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: ATCC 700721 / MGH 78578.
    2. "Biochemical characterization of the HpxO enzyme from Klebsiella pneumoniae, a novel FAD-dependent urate oxidase."
      O'Leary S.E., Hicks K.A., Ealick S.E., Begley T.P.
      Biochemistry 48:3033-3035(2009) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY.
      Strain: ATCC 700721 / MGH 78578.
    3. "Structural and mechanistic studies of HpxO, a novel flavin adenine dinucleotide-dependent urate oxidase from Klebsiella pneumoniae."
      Hicks K.A., O'Leary S.E., Begley T.P., Ealick S.E.
      Biochemistry 52:477-487(2013) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.97 ANGSTROMS) OF WILD-TYPE AND MUTANT GLN-204 IN COMPLEXES WITH URATE AND FAD, SUBUNIT, REACTION MECHANISM, MUTAGENESIS OF ARG-204 AND TYR-216.
      Strain: ATCC 700721 / MGH 78578.

    Entry informationi

    Entry nameiHPXO_KLEP7
    AccessioniPrimary (citable) accession number: A6T923
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: September 5, 2012
    Last sequence update: August 21, 2007
    Last modified: April 13, 2016
    This is version 50 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Complete proteome, Reference proteome

    Documents

    1. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    2. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    3. SIMILARITY comments
      Index of protein domains and families

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.