Reviewed, UniProtKB/Swiss-Prot Q1LCS4 (3HAO_RALME)

Names and origin

Protein names

Recommended name:
    3-hydroxyanthranilate 3,4-dioxygenase
    EC=1.13.11.6
Alternative name(s):
    3-hydroxyanthranilic acid dioxygenase
      Short name=HAD
    3-hydroxyanthranilate oxygenase
      Short name=3-HAO

Gene names

Ordered Locus Names:

Rmet_5193

Encoded on

Plasmid megaplasmid

Organism

Ralstonia metallidurans (strain CH34 / ATCC 43123 / DSM 2839) [Complete proteome] [HAMAP]

Taxonomic identifier

266264 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Betaproteobacteria › Burkholderiales › Burkholderiaceae › Cupriavidus

Protein attributes

Sequence length	174 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Catalyzes the oxidative ring opening of 3-hydroxyanthranilate to 2-amino-3-carboxymuconate semialdehyde, which spontaneously cyclizes to quinolinate.
Catalytic activity	3-hydroxyanthranilate + O(2) = 2-amino-3-carboxymuconate semialdehyde.
Cofactor	Binds 2 Fe(2+) ions per subunit.
Enzyme regulation	Inhibited by 4-chloro-3-hydroxyanthranilate. Mechanism of inactivation involves the oxidation of the catalytic active site Fe(2+) to the catalytically inactive Fe(3+) oxidation state, superoxide production, and formation of two disulfide bonds between Cys-125 and Cys-128, and Cys-162 and Cys-165. Enzyme can be reactivated under reducing conditions.
Subunit structure	Homodimer.
Sequence similarities	Belongs to the 3-HAO family.
Biophysicochemical properties	Kinetic parameters: KM=22.4 µM for 3-hydroxyanthranilate

Ontologies

Keywords
Ligand	Iron Metal-binding
Molecular function	Dioxygenase Oxidoreductase
Technical term	3D-structure Complete proteome Plasmid
Gene Ontology (GO)
Biological process	oxidation reduction Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	3-hydroxyanthranilate 3,4-dioxygenase activity Inferred from electronic annotation. Source: HAMAP ferrous iron binding Inferred from electronic annotation. Source: HAMAP
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 174

174

3-hydroxyanthranilate 3,4-dioxygenase

PRO_0000245477

Sites

Metal binding

51

1

Iron 1; catalytic

Metal binding

57

1

Iron 1; catalytic

Metal binding

95

1

Iron 1; catalytic

Metal binding

125

1

Iron 2

Metal binding

128

1

Iron 2

Metal binding

162

1

Iron 2

Metal binding

165

1

Iron 2

Binding site

47

1

Dioxygen

Binding site

57

1

Substrate

Binding site

99

1

Substrate

Binding site

110

1

Substrate

Experimental info

Mutagenesis

47

1

R → A: Increases KM for 3-hydroxyanthranilate 7-fold. Decreases activity 1000-fold

Mutagenesis

99

1

R → A: Increases KM for 3-hydroxyanthranilate 40-fold. Decreases activity 5000-fold

Mutagenesis

110

1

E → A: Decreases KM for 3-hydroxyanthranilate 2-fold. Decreases activity 2000-fold

Secondary structure

1

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174

Helix Strand Turn

Details...

Helix

10 – 16

7

Helix

17 – 20

4

Turn

23 – 25

3

Beta strand

27 – 33

7

Beta strand

35 – 41

7

Beta strand

50 – 52

3

Beta strand

57 – 64

8

Beta strand

66 – 72

7

Beta strand

75 – 81

7

Beta strand

85 – 89

5

Beta strand

95 – 99

5

Beta strand

105 – 111

7

Beta strand

119 – 124

6

Turn

126 – 128

3

Beta strand

131 – 137

7

Helix

142 – 145

4

Helix

147 – 154

8

Helix

157 – 160

4

Turn

163 – 165

3

Sequences

	Sequence		Length	Mass (Da)
	Q1LCS4-1 [UniParc]. Last modified May 30, 2006. Version 1. Checksum: 385F2E3DEB3947B8	FASTA	174	20,028
	10 20 30 40 50 60 MLTYGAPFNF PRWIDEHAHL LKPPVGNRQV WQDSDFIVTV VGGPNHRTDY HDDPLEEFFY 70 80 90 100 110 120 QLRGNAYLNL WVDGRRERAD LKEGDIFLLP PHVRHSPQRP EAGSACLVIE RQRPAGMLDG 130 140 150 160 170 FEWYCDACGH LVHRVEVQLK SIVTDLPPLF ESFYASEDKR RCPHCGQVHP GRAA

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Complete sequence of the megaplasmid of Ralstonia metallidurans CH34." Copeland A., Lucas S., Lapidus A., Barry K., Detter J.C., Glavina del Rio T., Hammon N., Israni S., Dalin E., Tice H., Martinez M., Goltsman E., Pitluck S., Schmutz J., Larimer F., Land M., Hauser L., Kyrpides N. , Kim E., Mergeay M., Benotmane M.A., Vallaeys T., Michaux A., Monchy S., Dunn J., McCorkle S., Taghavi S., van der Lelie D., Richardson P. Submitted (APR-2006) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
[2]	"The mechanism of inactivation of 3-hydroxyanthranilate-3,4-dioxygenase by 4-chloro-3-hydroxyanthranilate." Colabroy K.L., Zhai H., Li T., Ge Y., Zhang Y., Liu A., Ealick S.E., McLafferty F.W., Begley T.P. Biochemistry 44:7623-7631(2005) [PubMed: 15909977] [Abstract] Cited for: FUNCTION, ENZYME REGULATION, MASS SPECTROMETRY.
[3]	"Structural studies on 3-hydroxyanthranilate-3,4-dioxygenase: the catalytic mechanism of a complex oxidation involved in NAD biosynthesis." Zhang Y., Colabroy K.L., Begley T.P., Ealick S.E. Biochemistry 44:7632-7643(2005) [PubMed: 15909978] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF NATIVE PROTEIN AND COMPLEXES WITH SUBSTRATES OR INHIBITOR AND IRON, COFACTOR, KINETIC PARAMETERS, MUTAGENESIS OF ARG-47; ARG-99 AND GLU-110, SUBUNIT, REACTION MECHANISM.

Cross-references

Sequence databases

EMBLGenBankDDBJ

CP000353 Genomic DNA. Translation: ABF12052.1.

RefSeq

YP_587321.1.

3D structure databases

PDBMSD

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1YFU	X-ray	1.90	A/B	1-174	[»]
1YFW	X-ray	2.00	A/B	1-174	[»]
1YFX	X-ray	2.00	A/B	1-174	[»]
1YFY	X-ray	3.20	A/B	1-174	[»]

ModBase

Search...

Genome annotation databases

GeneID

4042054.

GenomeReviews

Gene locus Rmet_5193 in contig CP000353_GR.

KEGG

rme:Rmet_5193.

Organism-specific databases

CMR

Search...

Phylogenomic databases

HOGENOM

Q1LCS4.

Enzyme and pathway databases

BioCyc

RMET266264:RMET_5193-MON.

Family and domain databases

HAMAP

MF_00825.
[Tree]

InterPro

IPR010329. 3hydroanth_dOase.
[Graphical view]

Pfam

PF06052. 3-HAO. 1 hit.
[Graphical view]

TIGRFAMs

TIGR03037. anthran_nbaC. 1 hit.

ProtoNet

Search...

Entry information

Entry name

3HAO_RALME

Accession

Primary (citable) accession number: Q1LCS4

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 11, 2006
Last sequence update:	May 30, 2006
Last modified:	November 4, 2008
This is version 23 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)