AURF_STRTU
ID AURF_STRTU Reviewed; 336 AA.
AC Q70KH9;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=4-aminobenzoate N-oxygenase {ECO:0000305};
DE EC=1.14.99.68 {ECO:0000269|PubMed:15038705, ECO:0000269|PubMed:16927313, ECO:0000269|PubMed:18458342, ECO:0000269|PubMed:19731912, ECO:0000269|PubMed:20798054};
DE AltName: Full=N-oxygenase {ECO:0000303|PubMed:15038705};
DE AltName: Full=Non-heme di-iron N-oxygenase {ECO:0000305};
DE AltName: Full=p-aminobenzoate oxygenase {ECO:0000303|PubMed:16927313};
GN Name=aurF {ECO:0000303|PubMed:14700630};
OS Streptomyces thioluteus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=66431;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND PATHWAY.
RC STRAIN=HKI-227;
RX PubMed=14700630; DOI=10.1016/j.chembiol.2003.11.009;
RA He J., Hertweck C.;
RT "Iteration as programmed event during polyketide assembly; molecular
RT analysis of the aureothin biosynthesis gene cluster.";
RL Chem. Biol. 10:1225-1232(2003).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, DISRUPTION PHENOTYPE, AND PATHWAY.
RX PubMed=15038705; DOI=10.1021/ja039328t;
RA He J., Hertweck C.;
RT "Biosynthetic origin of the rare nitroaryl moiety of the polyketide
RT antibiotic aureothin: involvement of an unprecedented N-oxygenase.";
RL J. Am. Chem. Soc. 126:3694-3695(2004).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF GLU-101;
RP ASP-135; GLU-136; HIS-139; GLU-196; ASP-226; GLU-227 AND HIS-230.
RC STRAIN=ATCC 12310 / DSM 40027 / JCM 4087 / NBRC 13341 / NRRL B-1667 / NIHJ
RC 26A;
RX PubMed=16927313; DOI=10.1002/cbic.200600136;
RA Simurdiak M., Lee J., Zhao H.;
RT "A new class of arylamine oxygenases: evidence that p-aminobenzoate N-
RT oxygenase (AurF) is a di-iron enzyme and further mechanistic studies.";
RL ChemBioChem 7:1169-1172(2006).
RN [4]
RP COFACTOR.
RX PubMed=17718517; DOI=10.1021/bi701060g;
RA Krebs C., Matthews M.L., Jiang W., Bollinger J.M. Jr.;
RT "AurF from Streptomyces thioluteus and a possible new family of
RT manganese/iron oxygenases.";
RL Biochemistry 46:10413-10418(2007).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM, AND COFACTOR.
RX PubMed=19731912; DOI=10.1021/ja9064969;
RA Korboukh V.K., Li N., Barr E.W., Bollinger J.M. Jr., Krebs C.;
RT "A long-lived, substrate-hydroxylating peroxodiiron(III/III) intermediate
RT in the amine oxygenase, AurF, from Streptomyces thioluteus.";
RL J. Am. Chem. Soc. 131:13608-13609(2009).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM, AND COFACTOR.
RX PubMed=20798054; DOI=10.1073/pnas.1002785107;
RA Li N., Korboukh V.K., Krebs C., Bollinger J.M. Jr.;
RT "Four-electron oxidation of p-hydroxylaminobenzoate to p-nitrobenzoate by a
RT peroxodiferric complex in AurF from Streptomyces thioluteus.";
RL Proc. Natl. Acad. Sci. U.S.A. 107:15722-15727(2010).
RN [7] {ECO:0007744|PDB:2JCD}
RP X-RAY CRYSTALLOGRAPHY (2.11 ANGSTROMS) IN COMPLEX WITH MANGANESE, COFACTOR,
RP SUBUNIT, AND MUTAGENESIS OF ARG-96; THR-100; LEU-202; PHE-264 AND LEU-300.
RX PubMed=17765264; DOI=10.1016/j.jmb.2007.06.014;
RA Zocher G., Winkler R., Hertweck C., Schulz G.E.;
RT "Structure and action of the N-oxygenase AurF from Streptomyces
RT thioluteus.";
RL J. Mol. Biol. 373:65-74(2007).
RN [8] {ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI, ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEXES WITH IRON AND
RP 4-NITROBENZOIC ACID, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, COFACTOR, AND SUBUNIT.
RX PubMed=18458342; DOI=10.1073/pnas.0712073105;
RA Choi Y.S., Zhang H., Brunzelle J.S., Nair S.K., Zhao H.;
RT "In vitro reconstitution and crystal structure of p-aminobenzoate N-
RT oxygenase (AurF) involved in aureothin biosynthesis.";
RL Proc. Natl. Acad. Sci. U.S.A. 105:6858-6863(2008).
CC -!- FUNCTION: Involved in the biosynthesis of the polyketide antibiotic
CC aureothin (PubMed:14700630, PubMed:15038705). Catalyzes the oxidation
CC of p-aminobenzoate (pABA) to p-nitrobenzoate (pNBA), an unusual
CC polyketide synthase starter unit (PubMed:15038705, PubMed:16927313,
CC PubMed:20798054, PubMed:18458342). Reaction mechanism involves the
CC generation of a peroxodiiron(III/III) intermediate, which effects the
CC initial oxidation of p-aminobenzoate to p-hydroxylaminobenzoate (Ar-
CC NHOH) (PubMed:19731912, PubMed:20798054). Ar-NHOH is then probably
CC directly converted to the fully oxidized p-nitrobenzoate via a four-
CC electron N-oxidation, bypassing the formation of a nitroso compound
CC (PubMed:20798054). {ECO:0000269|PubMed:14700630,
CC ECO:0000269|PubMed:15038705, ECO:0000269|PubMed:16927313,
CC ECO:0000269|PubMed:18458342, ECO:0000269|PubMed:19731912,
CC ECO:0000269|PubMed:20798054}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-aminobenzoate + AH2 + 2 O2 = 4-nitrobenzoate + A + 2 H2O;
CC Xref=Rhea:RHEA:58888, ChEBI:CHEBI:13193, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17499, ChEBI:CHEBI:17836,
CC ChEBI:CHEBI:142863; EC=1.14.99.68;
CC Evidence={ECO:0000269|PubMed:15038705, ECO:0000269|PubMed:16927313,
CC ECO:0000269|PubMed:18458342, ECO:0000269|PubMed:19731912,
CC ECO:0000269|PubMed:20798054};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:58889;
CC Evidence={ECO:0000269|PubMed:15038705, ECO:0000269|PubMed:16927313,
CC ECO:0000269|PubMed:18458342, ECO:0000269|PubMed:20798054};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:16927313, ECO:0000269|PubMed:18458342,
CC ECO:0000269|PubMed:19731912, ECO:0000269|PubMed:20798054};
CC Note=Contains a nonheme dinuclear iron cluster that stabilizes a peroxo
CC intermediate (PubMed:16927313, PubMed:18458342, PubMed:19731912,
CC PubMed:20798054). Was originally suggested to contain a binuclear
CC manganese cluster or an heterodinuclear manganese/iron cluster
CC (PubMed:17765264, PubMed:17718517). {ECO:0000269|PubMed:16927313,
CC ECO:0000269|PubMed:17718517, ECO:0000269|PubMed:17765264,
CC ECO:0000269|PubMed:18458342, ECO:0000269|PubMed:19731912,
CC ECO:0000269|PubMed:20798054};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.24 uM for p-aminobenzoate {ECO:0000269|PubMed:18458342};
CC Note=kcat is 6.21 min(-1) with p-aminobenzoate as substrate.
CC {ECO:0000269|PubMed:18458342};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000269|PubMed:14700630,
CC ECO:0000269|PubMed:15038705}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:17765264,
CC ECO:0000269|PubMed:18458342}.
CC -!- INTERACTION:
CC Q70KH9; Q70KH9: aurF; NbExp=2; IntAct=EBI-15700476, EBI-15700476;
CC -!- DISRUPTION PHENOTYPE: Deletion of the gene abolishes both N-oxidation
CC activity and aureothin production. {ECO:0000269|PubMed:15038705}.
CC -!- SIMILARITY: Belongs to the AurF N-oxygenase family. {ECO:0000305}.
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DR EMBL; AJ575648; CAE02601.1; -; Genomic_DNA.
DR PDB; 2JCD; X-ray; 2.11 A; A/B=1-336.
DR PDB; 3CHH; X-ray; 2.00 A; A/B=1-336.
DR PDB; 3CHI; X-ray; 2.10 A; A/B=1-336.
DR PDB; 3CHT; X-ray; 2.00 A; A/B=1-336.
DR PDB; 3CHU; X-ray; 2.20 A; A/B=1-336.
DR PDBsum; 2JCD; -.
DR PDBsum; 3CHH; -.
DR PDBsum; 3CHI; -.
DR PDBsum; 3CHT; -.
DR PDBsum; 3CHU; -.
DR SMR; Q70KH9; -.
DR DIP; DIP-46089N; -.
DR KEGG; ag:CAE02601; -.
DR BioCyc; MetaCyc:MON-16310; -.
DR BRENDA; 1.14.99.68; 12297.
DR EvolutionaryTrace; Q70KH9; -.
DR GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR Gene3D; 1.10.620.20; -; 1.
DR InterPro; IPR025859; AurF.
DR InterPro; IPR012348; RNR-like.
DR Pfam; PF11583; AurF; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Iron; Manganese; Metal-binding; Oxidoreductase.
FT CHAIN 1..336
FT /note="4-aminobenzoate N-oxygenase"
FT /id="PRO_0000454383"
FT BINDING 93
FT /ligand="4-nitrobenzoate"
FT /ligand_id="ChEBI:CHEBI:142863"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHT"
FT BINDING 101
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 136
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 136
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 139
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 196
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 200
FT /ligand="4-nitrobenzoate"
FT /ligand_id="ChEBI:CHEBI:142863"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHT"
FT BINDING 223
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 227
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT BINDING 230
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:18458342,
FT ECO:0007744|PDB:3CHH, ECO:0007744|PDB:3CHI,
FT ECO:0007744|PDB:3CHT, ECO:0007744|PDB:3CHU"
FT MUTAGEN 96
FT /note="R->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:17765264"
FT MUTAGEN 100
FT /note="T->A: 3-fold increase in activity."
FT /evidence="ECO:0000269|PubMed:17765264"
FT MUTAGEN 100
FT /note="T->L: Retains 14% of activity."
FT /evidence="ECO:0000269|PubMed:17765264"
FT MUTAGEN 101
FT /note="E->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 135
FT /note="D->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 136
FT /note="E->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 139
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 196
FT /note="E->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 202
FT /note="L->F: 3.5-fold increase in activity."
FT /evidence="ECO:0000269|PubMed:17765264"
FT MUTAGEN 226
FT /note="D->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 227
FT /note="E->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 230
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:16927313"
FT MUTAGEN 264
FT /note="F->A: No change in activity."
FT /evidence="ECO:0000269|PubMed:17765264"
FT MUTAGEN 300
FT /note="L->W: Retains 80% of activity."
FT /evidence="ECO:0000269|PubMed:17765264"
FT HELIX 26..38
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 40..43
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 47..51
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 63..65
FT /evidence="ECO:0007829|PDB:3CHH"
FT TURN 67..70
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 72..75
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 79..103
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 105..113
FT /evidence="ECO:0007829|PDB:3CHH"
FT TURN 118..121
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 123..153
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 166..175
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 181..197
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 201..207
FT /evidence="ECO:0007829|PDB:3CHH"
FT STRAND 210..212
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 214..244
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 247..265
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 270..279
FT /evidence="ECO:0007829|PDB:3CHH"
FT HELIX 284..290
FT /evidence="ECO:0007829|PDB:3CHH"
FT TURN 295..298
FT /evidence="ECO:0007829|PDB:2JCD"
FT HELIX 305..313
FT /evidence="ECO:0007829|PDB:3CHH"
FT TURN 317..319
FT /evidence="ECO:0007829|PDB:3CHH"
SQ SEQUENCE 336 AA; 38083 MW; 9521375B4D5226C3 CRC64;
MREEQPHLAT TWAARGWVEE EGIGSATLGR LVRAWPRRAA VVNKADILDE WADYDTLVPD
YPLEIVPFAE HPLFLAAEPH QRQRVLTGMW IGYNERVIAT EQLIAEPAFD LVMHGVFPGS
DDPLIRKSVQ QAIVDESFHT YMHMLAIDRT RELRKISERP PQPELVTYRR LRRVLADMPE
QWERDIAVLV WGAVAETCIN ALLALLARDA TIQPMHSLIT TLHLRDETAH GSIVVEVVRE
LYARMNEQQR RALVRCLPIA LEAFAEQDLS ALLLELNAAG IRGAEEIVGD LRSTAGGTRL
VRDFSGARKM VEQLGLDDAV DFDFPERPDW SPHTPR
