XANA_EMEND
ID XANA_EMEND Reviewed; 370 AA.
AC Q4QZZ9;
DT 16-JAN-2019, integrated into UniProtKB/Swiss-Prot.
DT 19-JUL-2005, sequence version 1.
DT 03-AUG-2022, entry version 54.
DE RecName: Full=Alpha-ketoglutarate-dependent xanthine dioxygenase xanA {ECO:0000303|PubMed:15948966};
DE EC=1.14.11.48 {ECO:0000269|PubMed:15948966, ECO:0000269|PubMed:17429948, ECO:0000269|PubMed:18036331};
GN Name=xanA {ECO:0000303|PubMed:15948966};
OS Emericella nidulans (Aspergillus nidulans).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Nidulantes.
OX NCBI_TaxID=162425;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF ALA-167.
RC STRAIN=Glasgow wild type;
RX PubMed=15948966; DOI=10.1111/j.1365-2958.2005.04686.x;
RA Cultrone A., Scazzocchio C., Rochet M., Montero-Moran G., Drevet C.,
RA Fernandez-Martin R.;
RT "Convergent evolution of hydroxylation mechanisms in the fungal kingdom:
RT molybdenum cofactor-independent hydroxylation of xanthine via alpha-
RT ketoglutarate-dependent dioxygenases.";
RL Mol. Microbiol. 57:276-290(2005).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP ACTIVITY REGULATION, SUBSTRATE SPECIFICITY, PHOSPHORYLATION, AND
RP GLYCOSYLATION.
RX PubMed=17429948; DOI=10.1021/bi700065h;
RA Montero-Moran G.M., Li M., Rendon-Huerta E., Jourdan F., Lowe D.J.,
RA Stumpff-Kane A.W., Feig M., Scazzocchio C., Hausinger R.P.;
RT "Purification and characterization of the FeII- and alpha-ketoglutarate-
RT dependent xanthine hydroxylase from Aspergillus nidulans.";
RL Biochemistry 46:5293-5304(2007).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, SUBSTRATE
RP SPECIFICITY, AND MUTAGENESIS OF GLN-101; LYS-122; GLU-137; ASP-138;
RP HIS-149; ASP-151; HIS-340; GLN-356; CYS-357 AND ASN-358.
RX PubMed=18036331; DOI=10.1016/j.abb.2007.11.002;
RA Li M., Mueller T.A., Fraser B.A., Hausinger R.P.;
RT "Characterization of active site variants of xanthine hydroxylase from
RT Aspergillus nidulans.";
RL Arch. Biochem. Biophys. 470:44-53(2008).
RN [4]
RP SUBCELLULAR LOCATION.
RX PubMed=24970358; DOI=10.1016/j.fgb.2014.06.005;
RA Galanopoulou K., Scazzocchio C., Galinou M.E., Liu W., Borbolis F.,
RA Karachaliou M., Oestreicher N., Hatzinikolaou D.G., Diallinas G.,
RA Amillis S.;
RT "Purine utilization proteins in the Eurotiales: cellular
RT compartmentalization, phylogenetic conservation and divergence.";
RL Fungal Genet. Biol. 69:96-108(2014).
CC -!- FUNCTION: Alpha-ketoglutarate-dependent xanthine dioxygenase is a non-
CC heme mononuclear Fe(2+) enzyme that decarboxylates alpha-ketoglutarate
CC to succinate and CO(2) while hydroxylating xanthine to generate uric
CC acid (PubMed:15948966, PubMed:17429948, PubMed:18036331). Allows
CC xanthine utilization as a nitrogen source (PubMed:15948966). Whereas
CC xanA is highly specific for xanthine, alpha-ketoadipic acid can replace
CC alpha-ketoglutarate as a cosubstrate (PubMed:17429948). Exhibits
CC ferroxidase activity in the absence of substrates (PubMed:18036331).
CC {ECO:0000269|PubMed:15948966, ECO:0000269|PubMed:17429948,
CC ECO:0000269|PubMed:18036331}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + O2 + xanthine = CO2 + succinate + urate;
CC Xref=Rhea:RHEA:43120, ChEBI:CHEBI:15379, ChEBI:CHEBI:16526,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:17712, ChEBI:CHEBI:17775,
CC ChEBI:CHEBI:30031; EC=1.14.11.48;
CC Evidence={ECO:0000269|PubMed:15948966, ECO:0000269|PubMed:17429948,
CC ECO:0000269|PubMed:18036331};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:43121;
CC Evidence={ECO:0000269|PubMed:15948966, ECO:0000269|PubMed:17429948,
CC ECO:0000269|PubMed:18036331};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:17429948, ECO:0000269|PubMed:18036331};
CC Note=Binds 1 Fe(2+) ion per subunit. {ECO:0000250|UniProtKB:P37610};
CC -!- ACTIVITY REGULATION: Cu(2+) and Zn(2+) completely inhibit the xanthine
CC dioxygenase activity, whereas Co(2+), Mn(2+), and Ni(2+) partially
CC inhibit the activity. The inactive metal ions are presumed to compete
CC for the Fe(2+)-binding site (PubMed:17429948). N-oxalylglycine (NOG), a
CC known inhibitor of several Fe(2+)/alpha-ketoglutarate-dependent
CC dioxygenase family members, competes with alpha-ketoglutarate and
CC provides a Ki of 0.12 uM for inhibition (PubMed:17429948). 6,8-
CC dihydroxypurine acts as a slow-binding competitive inhibitor
CC (PubMed:18036331). The thiol-specific inhibitors 5,5'-dithiobis(2-
CC nitrobenzoic acid) (DTNB) and iodoacetamide, inhibit also the catalytic
CC activity (PubMed:18036331). {ECO:0000269|PubMed:17429948,
CC ECO:0000269|PubMed:18036331}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=31.1 uM for 2-oxoglutarate (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:17429948, ECO:0000269|PubMed:18036331};
CC KM=50.0 uM for 2-oxoglutarate (at 30 degrees Celsius)
CC {ECO:0000269|PubMed:15948966, ECO:0000269|PubMed:17429948};
CC KM=45.2 uM for xanthine (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:17429948, ECO:0000269|PubMed:18036331};
CC KM=46.0 uM for xanthine (at 30 degrees Celsius)
CC {ECO:0000269|PubMed:15948966, ECO:0000269|PubMed:17429948};
CC KM=0.16 mM for alpha-ketoadipic acid (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:17429948};
CC KM=0.4 mM for 9-methylxanthine (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:18036331};
CC pH dependence:
CC Optimum pH is 6.5 to 7.4. {ECO:0000269|PubMed:15948966,
CC ECO:0000269|PubMed:17429948};
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol {ECO:0000269|PubMed:24970358}.
CC -!- PTM: Glycosylated (PubMed:17429948). Is subject to both N- and O-linked
CC glycosylation (PubMed:17429948). {ECO:0000269|PubMed:17429948}.
CC -!- PTM: Phosphorylated. {ECO:0000269|PubMed:17429948}.
CC -!- SIMILARITY: Belongs to the TfdA dioxygenase family. {ECO:0000305}.
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DR EMBL; AJ877916; CAI47587.1; -; Genomic_DNA.
DR AlphaFoldDB; Q4QZZ9; -.
DR SMR; Q4QZZ9; -.
DR BRENDA; 1.14.11.48; 517.
DR GO; GO:0005829; C:cytosol; IEA:UniProtKB-SubCell.
DR GO; GO:0097641; F:alpha-ketoglutarate-dependent xanthine dioxygenase activity; IEA:RHEA.
DR GO; GO:0051213; F:dioxygenase activity; IDA:AspGD.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0034418; P:urate biosynthetic process; IMP:AspGD.
DR GO; GO:0009115; P:xanthine catabolic process; IDA:AspGD.
DR Gene3D; 3.60.130.10; -; 1.
DR InterPro; IPR042098; TauD-like_sf.
DR InterPro; IPR003819; TauD/TfdA-like.
DR Pfam; PF02668; TauD; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Dioxygenase; Glycoprotein; Iron; Metal-binding; Oxidoreductase;
KW Phosphoprotein.
FT CHAIN 1..370
FT /note="Alpha-ketoglutarate-dependent xanthine dioxygenase
FT xanA"
FT /id="PRO_0000446004"
FT BINDING 107
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P37610"
FT BINDING 149
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250|UniProtKB:P37610,
FT ECO:0000269|PubMed:18036331"
FT BINDING 151
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250|UniProtKB:P37610,
FT ECO:0000269|PubMed:18036331"
FT BINDING 195
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000250|UniProtKB:P37610"
FT BINDING 325
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000250|UniProtKB:P37610"
FT BINDING 340
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250|UniProtKB:P37610,
FT ECO:0000269|PubMed:18036331"
FT BINDING 352
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000250|UniProtKB:P37610"
FT MUTAGEN 101
FT /note="Q->A: Leads to elevated ferroxidase activity in the
FT absence of substrates."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 122
FT /note="K->A: Affects the binding of 2-oxoglutarate."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 137
FT /note="E->A: Exhibits relatively enhanced activity and
FT affects the inhibition by 6,8-dihydroxypurine."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 138
FT /note="D->A: Exhibits relatively enhanced activity and
FT affects the inhibition by 6,8-dihydroxypurine."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 149
FT /note="H->A: Impairs catalytic activity."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 151
FT /note="D->A: Impairs catalytic activity."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 167
FT /note="A->D: In xanA1; impairs catalytic activity."
FT /evidence="ECO:0000269|PubMed:15948966"
FT MUTAGEN 340
FT /note="H->A: Exhibits only 0.17% of the wild-type enzyme
FT activity."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 356
FT /note="Q->A: Leads to elevated ferroxidase activity in the
FT absence of substrates and affects the inhibition by 6,8-
FT dihydroxypurine."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 357
FT /note="C->A: Exhibits relatively enhanced activity,
FT resistance to thiol-specific inhibitors such as DTNB or
FT iodoacetamide, and elevated ferroxidase activity in the
FT absence of substrates."
FT /evidence="ECO:0000269|PubMed:18036331"
FT MUTAGEN 358
FT /note="N->A: Exhibits a 23-fold decrease in kcat/Km and
FT affects the inhibition by 6,8-dihydroxypurine."
FT /evidence="ECO:0000269|PubMed:18036331"
SQ SEQUENCE 370 AA; 41305 MW; 5E3A746FDBFAC33A CRC64;
MPAITVKPLT PPAGSAIDFG AVITDVDLEH LTDGDFSTIR SALYTHLVVV LKNQHQLTPK
AQYELTRRFD PSATQYGHGK TLDAKRSILH PDLKTIPHQP QVQVIGHGFI DSYEGLENIT
LKHPHHRTFH RDPIPQEDDY DSTRFYRWHI DAALYGLNPP IVTTLLAVKV PGGRRQTVRY
DDGSGETMDV PLGTTAFASG ERMFELLSEE DKEFALSSRV EYAPHPYIWM SPARSLPTGL
GLHSDDLELP LSELPPIDES AIQILPMVWK NPATGKPALQ IHPSAVRKIH CGDGTVIDDL
KKVREIAYKL QRPAISPQYV YAHDWEEGDL VLFHNRGVLH SVVGAFGEGE VRLFRQCNLA
AGEGVVPYRE
