P00438 (PHHY_PSEFL) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
p-hydroxybenzoate hydroxylase
EC=1.14.13.2

Alternative name(s):
4-hydroxybenzoate 3-monooxygenase

Gene names

Name:

pobA

Organism

Pseudomonas fluorescens

Taxonomic identifier

294 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Pseudomonadales › Pseudomonadaceae › Pseudomonas › Pseudomonas fluorescens group

Protein attributes

Sequence length	394 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Catalytic activity	4-hydroxybenzoate + NADPH + O2 = protocatechuate + NADP+ + H2O.
Cofactor	FAD.
Pathway	Aromatic compound metabolism; benzoate degradation via hydroxylation; 3,4-dihydroxybenzoate from benzoate: step 2/2.
Subunit structure	Homodimer.

Ontologies

Keywords
Biological process	Aromatic hydrocarbons catabolism
Ligand	FAD Flavoprotein NADP
Molecular function	Monooxygenase Oxidoreductase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological_process	benzoate catabolic process via hydroxylation Inferred from electronic annotation. Source: UniProtKB-UniPathway
Molecular_function	4-hydroxybenzoate 3-monooxygenase activity Inferred from electronic annotation. Source: EC flavin adenine dinucleotide binding Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 394

394

p-hydroxybenzoate hydroxylase

PRO_0000058382

Regions

Nucleotide binding

4 – 33

30

FAD Potential

Nucleotide binding

276 – 286

11

FAD Potential

Experimental info

Sequence conflict

344

1

W → Y AA sequence Ref.2

Secondary structure

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394

Helix Strand Turn

Details...

Beta strand

4 – 8

5

Helix

12 – 23

12

Beta strand

28 – 31

4

Helix

36 – 40

5

Beta strand

47 – 49

3

Helix

50 – 58

9

Helix

63 – 68

6

Beta strand

70 – 79

10

Beta strand

82 – 87

6

Helix

88 – 91

4

Beta strand

97 – 99

3

Helix

102 – 116

15

Beta strand

119 – 121

3

Beta strand

125 – 130

6

Beta strand

134 – 136

3

Beta strand

138 – 143

6

Beta strand

146 – 151

6

Beta strand

153 – 157

5

Helix

164 – 167

4

Helix

171 – 173

3

Beta strand

175 – 192

18

Beta strand

195 – 198

4

Beta strand

200 – 203

4

Beta strand

209 – 215

7

Beta strand

218 – 225

8

Helix

231 – 233

3

Helix

236 – 246

11

Helix

249 – 254

6

Beta strand

260 – 278

19

Beta strand

281 – 283

3

Helix

285 – 287

3

Helix

293 – 295

3

Helix

298 – 318

21

Helix

322 – 327

6

Helix

328 – 350

23

Helix

358 – 371

14

Helix

375 – 386

12

Sequences

	Sequence		Length	Mass (Da)
	P00438 [UniParc]. Last modified April 1, 1993. Version 2. Checksum: D29599224AC81E00	FASTA	394	44,322
	10 20 30 40 50 60 MKTQVAIIGA GPSGLLLGQL LHKAGIDNVI LERQTPDYVL GRIRAGVLEQ GMVDLLREAG 70 80 90 100 110 120 VDRRMARDGL VHEGVEIAFA GQRRRIDLKR LSGGKTVTVY GQTEVTRDLM EAREACGATT 130 140 150 160 170 180 VYQAAEVRLH DLQGERPYVT FERDGERLRL DCDYIAGCDG FHGISRQSIP AERLKVFERV 190 200 210 220 230 240 YPFGWLGLLA DTPPVSHELI YANHPRGFAL CSQRSATRSR YYVQVPLTEK VEDWSDERFW 250 260 270 280 290 300 TELKARLPAE VAEKLVTGPS LEKSIAPLRS FVVEPMQHGR LFLAGDAAHI VPPTGAKGLN 310 320 330 340 350 360 LAASDVSTLY RLLLKAYREG RGELLERYSA ICLRRIWKAE RFSWWMTSVL HRFPDTDAFS 370 380 390 QRIQQTELEY YLGSEAGLAT IAENYVGLPY EEIE

References

[1]	"Substitution of Arg214 at the substrate-binding site of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens." van Berkel W., Westphal A., Eschrich K., Eppink M., de Kok A. Eur. J. Biochem. 210:411-419(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"Primary structure of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens." Weijer W.J., Hofsteenge J., Vereijken J.M., Jekel P.A., Beintema J.J. Biochim. Biophys. Acta 704:385-388(1982) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE.
[3]	"Primary and tertiary structure studies of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens. Isolation and alignment of the CNBr peptides; interactions of the protein with flavin adenine dinucleotide." Hofsteenge J., Vereijken J.M., Weijer W.J., Beintema J.J., Wierenga R.K., Drenth J. Eur. J. Biochem. 113:141-150(1980) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 111-138 AND 270-280.
[4]	"The amino-acid sequence of the three smallest CNBr peptides from p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens." Vereijken J.M., Hofsteenge J., Bak H.J., Beintema J.J. Eur. J. Biochem. 113:151-157(1980) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-52; 53-65 AND 66-110.
[5]	"p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens. 1. Completion of the elucidation of the primary structure." Hofsteenge J., Weijer W.J., Jekel P.A., Beintema J.J. Eur. J. Biochem. 133:91-108(1983) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF CNBR PEPTIDES AND TERTIARY STRUCTURE.
[6]	"p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens. 2. Fitting of the amino-acid sequence to the tertiary structure." Weijer W.J., Hofsteenge J., Beintema J.J., Wierenga R.K., Drenth J. Eur. J. Biochem. 133:109-118(1983) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF CNBR PEPTIDES, FAD BINDING SITE.
[7]	"Crystal structure of p-hydroxybenzoate hydroxylase." Wierenga R.K., de Jong R.J., Kalk K.H., Hol W.G.J., Drenth J. J. Mol. Biol. 131:55-73(1979) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS).
[8]	"Crystal structure of p-hydroxybenzoate hydroxylase complexed with its reaction product 3,4-dihydroxybenzoate." Schreuder H.A., van der Laan J.M., Hol W.G.J., Drenth J. J. Mol. Biol. 199:637-648(1988) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS).
[9]	"Crystal structure of the reduced form of p-hydroxybenzoate hydroxylase refined at 2.3-A resolution." Schreuder H.A., van der Laan J.M., Swarte M.B.A., Kalk K.H., Hol W.G.J., Drenth J. Proteins 14:178-190(1992) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF REDUCED FORM.
[10]	"Structure and function of mutant Arg44Lys of 4-hydroxybenzoate hydroxylase implications for NADPH binding." Eppink M.H., Schreuder H.A., van Berkel W.J. Eur. J. Biochem. 231:157-165(1995) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT LYS-44.
[11]	"Lys42 and Ser42 variants of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens reveal that Arg42 is essential for NADPH binding." Eppink M.H., Schreuder H.A., van Berkel W.J. Eur. J. Biochem. 253:194-201(1998) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT LYS-42 AND SER-42.
[12]	"Phe161 and Arg166 variants of p-hydroxybenzoate hydroxylase. Implications for NADPH recognition and structural stability." Eppink M.H., Bunthol C., Schreuder H.A., van Berkel W.J. FEBS Lett. 443:251-255(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF VARIANTS.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

X68438 Genomic DNA. Translation: CAA48483.1.

PIR

WHPSBF. A90643.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1BF3	X-ray	2.20	A	1-394	[»]
1BGJ	X-ray	3.00	A	1-394	[»]
1BGN	X-ray	2.00	A	1-394	[»]
1BKW	X-ray	2.20	A	1-394	[»]
1CC4	X-ray	2.00	A	1-394	[»]
1CC6	X-ray	2.20	A	1-394	[»]
1CJ2	X-ray	2.80	A	1-391	[»]
1CJ3	X-ray	2.50	A	1-392	[»]
1CJ4	X-ray	2.40	A	1-392	[»]
1PBB	X-ray	2.50	A	1-394	[»]
1PBC	X-ray	2.80	A	1-394	[»]
1PBD	X-ray	2.30	A	1-394	[»]
1PBE	X-ray	1.90	A	1-394	[»]
1PBF	X-ray	2.70	A	1-394	[»]
1PDH	X-ray	2.10	A	1-394	[»]
1PHH	X-ray	2.30	A	1-394	[»]
2PHH	X-ray	2.70	A	1-394	[»]

ProteinModelPortal

P00438.

SMR

P00438. Positions 1-394.

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Enzyme and pathway databases

BioCyc

MetaCyc:MONOMER-11534.

UniPathway

UPA00156; UER00257.

Family and domain databases

InterPro

IPR012733. HB_mOase.
IPR002938. mOase_FAD-bd.
IPR003042. Rng_hydrolase-like.
[Graphical view]

Pfam

PF01494. FAD_binding_3. 1 hit.
[Graphical view]

PRINTS

PR00420. RNGMNOXGNASE.

TIGRFAMs

TIGR02360. pbenz_hydroxyl. 1 hit.

ProtoNet

Search...

Other

BindingDB

P00438.

ChEMBL

CHEMBL3675.

DrugBank

DB00233. Aminosalicylic Acid.

EvolutionaryTrace

P00438.

Entry information

Entry name

PHHY_PSEFL

Accession

Primary (citable) accession number: P00438

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 21, 1986
Last sequence update:	April 1, 1993
Last modified:	April 3, 2013
This is version 97 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program