HPXO_KLEP7
ID HPXO_KLEP7 Reviewed; 384 AA.
AC A6T923;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 21-AUG-2007, sequence version 1.
DT 03-AUG-2022, entry version 77.
DE RecName: Full=FAD-dependent urate hydroxylase {ECO:0000305|PubMed:19260710};
DE EC=1.14.13.113 {ECO:0000269|PubMed:19260710};
DE AltName: Full=FAD-dependent urate oxidase {ECO:0000303|PubMed:19260710};
GN Name=hpxO {ECO:0000303|PubMed:19260710}; OrderedLocusNames=KPN78578_16330;
GN ORFNames=KPN_01663;
OS Klebsiella pneumoniae subsp. pneumoniae (strain ATCC 700721 / MGH 78578).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Klebsiella/Raoultella group; Klebsiella.
OX NCBI_TaxID=272620;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700721 / MGH 78578;
RG The Klebsiella pneumonia Genome Sequencing Project;
RA McClelland M., Sanderson E.K., Spieth J., Clifton W.S., Latreille P.,
RA Sabo A., Pepin K., Bhonagiri V., Porwollik S., Ali J., Wilson R.K.;
RL Submitted (SEP-2006) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP PATHWAY.
RC STRAIN=ATCC 700721 / MGH 78578;
RX PubMed=19260710; DOI=10.1021/bi900160b;
RA O'Leary S.E., Hicks K.A., Ealick S.E., Begley T.P.;
RT "Biochemical characterization of the HpxO enzyme from Klebsiella
RT pneumoniae, a novel FAD-dependent urate oxidase.";
RL Biochemistry 48:3033-3035(2009).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.97 ANGSTROMS) OF WILD-TYPE AND MUTANT GLN-204 IN
RP COMPLEXES WITH URATE AND FAD, SUBUNIT, REACTION MECHANISM, AND MUTAGENESIS
RP OF ARG-204 AND TYR-216.
RC STRAIN=ATCC 700721 / MGH 78578;
RX PubMed=23259842; DOI=10.1021/bi301262p;
RA Hicks K.A., O'Leary S.E., Begley T.P., Ealick S.E.;
RT "Structural and mechanistic studies of HpxO, a novel flavin adenine
RT dinucleotide-dependent urate oxidase from Klebsiella pneumoniae.";
RL Biochemistry 52:477-487(2013).
CC -!- FUNCTION: Catalyzes the hydroxylation of urate to 5-hydroxyisourate
CC (HIU). {ECO:0000269|PubMed:19260710}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + NADH + O2 + urate = 5-hydroxyisourate + H2O + NAD(+);
CC Xref=Rhea:RHEA:27329, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17775, ChEBI:CHEBI:18072,
CC ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; EC=1.14.13.113;
CC Evidence={ECO:0000269|PubMed:19260710};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:19260710};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=42 uM for urate {ECO:0000269|PubMed:19260710};
CC Note=kcat is 42 sec(-1). {ECO:0000269|PubMed:19260710};
CC -!- PATHWAY: Purine metabolism; urate degradation.
CC {ECO:0000269|PubMed:19260710}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:23259842}.
CC -!- SIMILARITY: Belongs to the FAD-dependent urate hydroxylase family.
CC {ECO:0000305}.
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DR EMBL; CP000647; ABR77094.1; -; Genomic_DNA.
DR RefSeq; WP_002904783.1; NC_009648.1.
DR PDB; 3RP6; X-ray; 2.20 A; A=1-384.
DR PDB; 3RP7; X-ray; 2.04 A; A=1-384.
DR PDB; 3RP8; X-ray; 1.97 A; A=1-384.
DR PDBsum; 3RP6; -.
DR PDBsum; 3RP7; -.
DR PDBsum; 3RP8; -.
DR AlphaFoldDB; A6T923; -.
DR SMR; A6T923; -.
DR STRING; 272620.KPN_01663; -.
DR EnsemblBacteria; ABR77094; ABR77094; KPN_01663.
DR KEGG; kpn:KPN_01663; -.
DR HOGENOM; CLU_009665_19_5_6; -.
DR OMA; RWMLGYD; -.
DR BRENDA; 1.14.13.113; 2814.
DR UniPathway; UPA00394; -.
DR Proteomes; UP000000265; Chromosome.
DR GO; GO:0071949; F:FAD binding; IDA:UniProtKB.
DR GO; GO:0102099; F:FAD-dependent urate hydroxylase activity; IEA:UniProtKB-EC.
DR GO; GO:0016709; F: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; IDA:UniProtKB.
DR GO; GO:0004846; F:urate oxidase activity; IDA:CACAO.
DR GO; GO:0006144; P:purine nucleobase metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0019628; P:urate catabolic process; IDA:UniProtKB.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR002938; FAD-bd.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR Pfam; PF01494; FAD_binding_3; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; FAD; Flavoprotein; Monooxygenase; NAD; Oxidoreductase;
KW Purine metabolism; Reference proteome.
FT CHAIN 1..384
FT /note="FAD-dependent urate hydroxylase"
FT /id="PRO_0000418845"
FT BINDING 11
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP6, ECO:0007744|PDB:3RP7,
FT ECO:0007744|PDB:3RP8"
FT BINDING 30..31
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP6, ECO:0007744|PDB:3RP7,
FT ECO:0007744|PDB:3RP8"
FT BINDING 43
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP6, ECO:0007744|PDB:3RP7,
FT ECO:0007744|PDB:3RP8"
FT BINDING 125
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP6, ECO:0007744|PDB:3RP7,
FT ECO:0007744|PDB:3RP8"
FT BINDING 178
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP7"
FT BINDING 204
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP7"
FT BINDING 216..218
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP7"
FT BINDING 285
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP6, ECO:0007744|PDB:3RP7,
FT ECO:0007744|PDB:3RP8"
FT BINDING 295..299
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:23259842,
FT ECO:0007744|PDB:3RP6, ECO:0007744|PDB:3RP7,
FT ECO:0007744|PDB:3RP8"
FT SITE 204
FT /note="Involved in substrate activation for the transfer of
FT oxygen from the flavin hydroperoxide"
FT /evidence="ECO:0000305|PubMed:23259842"
FT MUTAGEN 204
FT /note="R->Q: 160-fold decrease in catalytic activity.
FT Results in the uncoupling of the NADH oxidation and urate
FT hydroxylation reactions."
FT /evidence="ECO:0000269|PubMed:23259842"
FT MUTAGEN 216
FT /note="Y->F: 5-fold decrease in catalytic activity. 2-fold
FT decrease in affinity for urate and increase in affinity for
FT NADH."
FT /evidence="ECO:0000269|PubMed:23259842"
FT STRAND 2..6
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 10..21
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 25..33
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 41..44
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 46..54
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 58..64
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 70..75
FT /evidence="ECO:0007829|PDB:3RP8"
FT TURN 76..78
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 81..86
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 88..94
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 99..102
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 103..114
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 116..118
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 119..122
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 125..131
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 134..139
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 144..152
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 161..165
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 172..184
FT /evidence="ECO:0007829|PDB:3RP8"
FT TURN 187..189
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 194..200
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 203..210
FT /evidence="ECO:0007829|PDB:3RP8"
FT TURN 211..213
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 214..222
FT /evidence="ECO:0007829|PDB:3RP8"
FT TURN 231..233
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 234..241
FT /evidence="ECO:0007829|PDB:3RP8"
FT TURN 242..244
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 247..255
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 258..260
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 262..268
FT /evidence="ECO:0007829|PDB:3RP8"
FT STRAND 280..282
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 284..286
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 292..294
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 297..313
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 318..347
FT /evidence="ECO:0007829|PDB:3RP8"
FT TURN 348..351
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 352..363
FT /evidence="ECO:0007829|PDB:3RP8"
FT HELIX 368..379
FT /evidence="ECO:0007829|PDB:3RP8"
SQ SEQUENCE 384 AA; 42107 MW; 4B656520269C3535 CRC64;
MKAIVIGAGI GGLSAAVALK QSGIDCDVYE AVKEIKPVGA AISVWPNGVK CMAHLGMGDI
METFGGPLRR MAYRDFRSGE NMTQFSLAPL IERTGSRPCP VSRAELQREM LDYWGRDSVQ
FGKRVTRCEE DADGVTVWFT DGSSASGDLL IAADGSHSAL RPWVLGFTPQ RRYAGYVNWN
GLVEIDEALA PGDQWTTFVG EGKRVSLMPV SAGRFYFFFD VPLPAGLAED RDTLRADLSR
YFAGWAPPVQ KLIAALDPQT TNRIEIHDIE PFSRLVRGRV ALLGDAGHST TPDIGQGGCA
AMEDAVVLGA VFRQTRDIAA ALREYEAQRC DRVRDLVLKA RKRCDITHGK DMQLTEAWYQ
ELREETGERI INGMCDTILS GPLG
