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Protein

p-hydroxybenzoate hydroxylase

Gene

pobA

Organism
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the incorporation of an atom of dioxygen into p-hydroxybenzoate (p-OHB) to form 3,4-dihydroxybenzoate (3,4DOHB). The reaction occurs in two parts: reduction of the flavin adenine dinucleotide (FAD) in the enzyme by reduced nicotinamide adenine dinucleotide phosphate (NADPH) in response to binding p-hydroxybenzoate to the enzyme and oxidation of reduced FAD with oxygen to form a hydroperoxide, which then oxygenates p-hydroxybenzoate.7 Publications

Catalytic activityi

4-hydroxybenzoate + NADPH + O2 = protocatechuate + NADP+ + H2O.1 Publication

Cofactori

FAD6 PublicationsNote: Binds 1 FAD per subunit.5 Publications

Pathwayi: benzoate degradation via hydroxylation

This protein is involved in step 2 of the subpathway that synthesizes 3,4-dihydroxybenzoate from benzoate.Curated
Proteins known to be involved in the 2 steps of the subpathway in this organism are:
  1. no protein annotated in this organism
  2. p-hydroxybenzoate hydroxylase (pobA)
This subpathway is part of the pathway benzoate degradation via hydroxylation, which is itself part of Aromatic compound metabolism.
View all proteins of this organism that are known to be involved in the subpathway that synthesizes 3,4-dihydroxybenzoate from benzoate, the pathway benzoate degradation via hydroxylation and in Aromatic compound metabolism.

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei13 – 131FAD6 Publications
Binding sitei32 – 321FAD6 Publications
Binding sitei102 – 1021FAD6 Publications
Binding sitei201 – 2011Substrate6 Publications
Sitei201 – 2011Important for catalytic activity1 Publication
Binding sitei222 – 2221Substrate6 Publications
Binding sitei286 – 2861FAD5 Publications
Binding sitei293 – 2931Substrate; via carbonyl oxygen6 Publications
Sitei385 – 3851Important for catalytic activity1 Publication

Regions

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Nucleotide bindingi42 – 476FAD6 Publications
Nucleotide bindingi299 – 3002FAD6 Publications

GO - Molecular functioni

  • 4-hydroxybenzoate 3-monooxygenase activity Source: UniProtKB
  • FAD binding Source: UniProtKB
  • flavin adenine dinucleotide binding Source: UniProtKB
  • monooxygenase activity Source: GO_Central

GO - Biological processi

Complete GO annotation...

Keywords - Molecular functioni

Monooxygenase, Oxidoreductase

Keywords - Biological processi

Aromatic hydrocarbons catabolism

Keywords - Ligandi

FAD, Flavoprotein, NADP

Enzyme and pathway databases

BRENDAi1.14.13.2. 5087.
UniPathwayiUPA00156; UER00257.

Names & Taxonomyi

Protein namesi
Recommended name:
p-hydroxybenzoate hydroxylase1 Publication (EC:1.14.13.21 Publication)
Short name:
PHBH1 Publication
Alternative name(s):
4-hydroxybenzoate 3-monooxygenaseCurated
Gene namesi
Name:pobA1 Publication
Ordered Locus Names:PA0247
OrganismiPseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Taxonomic identifieri208964 [NCBI]
Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaPseudomonadalesPseudomonadaceaePseudomonas
Proteomesi
  • UP000002438 Componenti: Chromosome

Organism-specific databases

PseudoCAPiPA0247.

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi45 – 451A → G: The positions of the substrate and the flavin are not altered. 1 Publication
Mutagenesisi201 – 2011Y → F: Reduction of hydroxylase activity. 2 Publications
Mutagenesisi220 – 2201R → Q: Lower affinity for p-OHB than the wild-type. 1 Publication
Mutagenesisi300 – 3001N → D: The side chain of Asp300 moves away from the flavin, disrupting the interactions of the carboxamide group with the flavin O(2) atom, and the alpha-helix H10 that begins at residue 297 is displaced, altering its dipole interactions with the flavin ring. 1 Publication
Mutagenesisi385 – 3851Y → F: The positions of the substrate and the flavin are not altered. 2 Publications

Chemistry

DrugBankiDB03147. Flavin adenine dinucleotide.

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 394394p-hydroxybenzoate hydroxylasePRO_0000058381Add
BLAST

Proteomic databases

PaxDbiP20586.

Interactioni

Subunit structurei

Homodimer.6 Publications

Protein-protein interaction databases

STRINGi208964.PA0247.

Structurei

Secondary structure

1
394
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi4 – 85Combined sources
Helixi12 – 2413Combined sources
Beta strandi28 – 314Combined sources
Helixi36 – 405Combined sources
Beta strandi47 – 493Combined sources
Helixi50 – 589Combined sources
Helixi63 – 686Combined sources
Beta strandi70 – 734Combined sources
Beta strandi75 – 795Combined sources
Beta strandi82 – 865Combined sources
Helixi88 – 925Combined sources
Beta strandi97 – 993Combined sources
Helixi102 – 11514Combined sources
Beta strandi119 – 1235Combined sources
Beta strandi125 – 1306Combined sources
Beta strandi134 – 1363Combined sources
Beta strandi138 – 1436Combined sources
Beta strandi146 – 1516Combined sources
Beta strandi153 – 1575Combined sources
Helixi166 – 1683Combined sources
Helixi171 – 1733Combined sources
Beta strandi175 – 19218Combined sources
Beta strandi195 – 1984Combined sources
Beta strandi200 – 2023Combined sources
Beta strandi209 – 2157Combined sources
Beta strandi218 – 2258Combined sources
Helixi231 – 2333Combined sources
Helixi236 – 24510Combined sources
Helixi249 – 2546Combined sources
Beta strandi260 – 27415Combined sources
Beta strandi276 – 2783Combined sources
Beta strandi281 – 2833Combined sources
Helixi285 – 2873Combined sources
Helixi293 – 2953Combined sources
Helixi298 – 31922Combined sources
Helixi322 – 3276Combined sources
Helixi328 – 35023Combined sources
Helixi358 – 37316Combined sources
Helixi375 – 38511Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1D7LX-ray2.20A1-394[»]
1DOBX-ray2.00A1-394[»]
1DOCX-ray2.00A1-394[»]
1DODX-ray2.10A1-394[»]
1DOEX-ray2.30A1-394[»]
1IUSX-ray2.20A1-394[»]
1IUTX-ray2.00A1-394[»]
1IUUX-ray2.00A1-394[»]
1IUVX-ray2.50A1-394[»]
1IUWX-ray2.00A1-394[»]
1IUXX-ray2.00A1-394[»]
1K0IX-ray1.80A1-394[»]
1K0JX-ray2.20A1-394[»]
1K0LX-ray2.00A1-394[»]
1PXAX-ray2.30A1-394[»]
1PXBX-ray2.30A1-394[»]
1PXCX-ray2.10A1-394[»]
1YKJX-ray2.00A/B1-394[»]
ProteinModelPortaliP20586.
SMRiP20586. Positions 1-394.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiP20586.

Family & Domainsi

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni212 – 2143Substrate binding6 Publications

Sequence similaritiesi

Belongs to the aromatic-ring hydroxylase family.Curated

Phylogenomic databases

eggNOGiENOG41072AC. Bacteria.
COG0654. LUCA.
HOGENOMiHOG000221233.
InParanoidiP20586.
KOiK00481.
OMAiVEPMQYG.
OrthoDBiEOG6PS5RW.
PhylomeDBiP20586.

Family and domain databases

Gene3Di3.50.50.60. 1 hit.
InterProiIPR002938. FAD-bd.
IPR023753. FAD/NAD-binding_dom.
IPR012733. HB_mOase.
[Graphical view]
PfamiPF01494. FAD_binding_3. 1 hit.
[Graphical view]
SUPFAMiSSF51905. SSF51905. 2 hits.
TIGRFAMsiTIGR02360. pbenz_hydroxyl. 1 hit.

Sequencei

Sequence statusi: Complete.

P20586-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MKTQVAIIGA GPSGLLLGQL LHKAGIDNVI LERQTPDYVL GRIRAGVLEQ
60 70 80 90 100
GMVDLLREAG VDRRMARDGL VHEGVEIAFA GQRRRIDLKR LSGGKTVTVY
110 120 130 140 150
GQTEVTRDLM EAREACGATT VYQAAEVRLH DLQGERPYVT FERDGERLRL
160 170 180 190 200
DCDYIAGCDG FHGISRQSIP AERLKVFERV YPFGWLGLLA DTPPVSHELI
210 220 230 240 250
YANHPRGFAL CSQRSATRSR YYVQVPLSEK VEDWSDERFW TELKARLPSE
260 270 280 290 300
VAEKLVTGPS LEKSIAPLRS FVVEPMQHGR LFLAGDAAHI VPPTGAKGLN
310 320 330 340 350
LAASDVSTLY RLLLKAYREG RGELLERYSA ICLRRIWKAE RFSWWMTSVL
360 370 380 390
HRFPDTDAFS QRIQQTELEY YLGSEAGLAT IAENYVGLPY EEIE
Length:394
Mass (Da):44,324
Last modified:February 1, 1991 - v1
Checksum:i1E7232854D9EC792
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
M23173 Genomic DNA. Translation: AAA88455.1.
AE004091 Genomic DNA. Translation: AAG03636.1.
PIRiJT0384. WHPSBA.
RefSeqiNP_248938.1. NC_002516.2.
WP_003112685.1. NZ_ASJY01000056.1.

Genome annotation databases

EnsemblBacteriaiAAG03636; AAG03636; PA0247.
GeneIDi882128.
KEGGipae:PA0247.
PATRICi19834710. VBIPseAer58763_0257.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
M23173 Genomic DNA. Translation: AAA88455.1.
AE004091 Genomic DNA. Translation: AAG03636.1.
PIRiJT0384. WHPSBA.
RefSeqiNP_248938.1. NC_002516.2.
WP_003112685.1. NZ_ASJY01000056.1.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1D7LX-ray2.20A1-394[»]
1DOBX-ray2.00A1-394[»]
1DOCX-ray2.00A1-394[»]
1DODX-ray2.10A1-394[»]
1DOEX-ray2.30A1-394[»]
1IUSX-ray2.20A1-394[»]
1IUTX-ray2.00A1-394[»]
1IUUX-ray2.00A1-394[»]
1IUVX-ray2.50A1-394[»]
1IUWX-ray2.00A1-394[»]
1IUXX-ray2.00A1-394[»]
1K0IX-ray1.80A1-394[»]
1K0JX-ray2.20A1-394[»]
1K0LX-ray2.00A1-394[»]
1PXAX-ray2.30A1-394[»]
1PXBX-ray2.30A1-394[»]
1PXCX-ray2.10A1-394[»]
1YKJX-ray2.00A/B1-394[»]
ProteinModelPortaliP20586.
SMRiP20586. Positions 1-394.
ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

STRINGi208964.PA0247.

Chemistry

DrugBankiDB03147. Flavin adenine dinucleotide.

Proteomic databases

PaxDbiP20586.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaiAAG03636; AAG03636; PA0247.
GeneIDi882128.
KEGGipae:PA0247.
PATRICi19834710. VBIPseAer58763_0257.

Organism-specific databases

PseudoCAPiPA0247.

Phylogenomic databases

eggNOGiENOG41072AC. Bacteria.
COG0654. LUCA.
HOGENOMiHOG000221233.
InParanoidiP20586.
KOiK00481.
OMAiVEPMQYG.
OrthoDBiEOG6PS5RW.
PhylomeDBiP20586.

Enzyme and pathway databases

UniPathwayiUPA00156; UER00257.
BRENDAi1.14.13.2. 5087.

Miscellaneous databases

EvolutionaryTraceiP20586.

Family and domain databases

Gene3Di3.50.50.60. 1 hit.
InterProiIPR002938. FAD-bd.
IPR023753. FAD/NAD-binding_dom.
IPR012733. HB_mOase.
[Graphical view]
PfamiPF01494. FAD_binding_3. 1 hit.
[Graphical view]
SUPFAMiSSF51905. SSF51905. 2 hits.
TIGRFAMsiTIGR02360. pbenz_hydroxyl. 1 hit.
ProtoNetiSearch...

Publicationsi

« Hide 'large scale' publications
  1. "Sequence and organization of pobA, the gene coding for p-hydroxybenzoate hydroxylase, an inducible enzyme from Pseudomonas aeruginosa."
    Entsch B., Nan Y., Weaich K., Scott K.F.
    Gene 71:279-291(1988) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
  2. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
    Strain: ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228.
  3. "Catalytic function of tyrosine residues in para-hydroxybenzoate hydroxylase as determined by the study of site-directed mutants."
    Entsch B., Palfey B.A., Ballou D.P., Massey V.
    J. Biol. Chem. 266:17341-17349(1991) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF TYR-201 AND TYR-385, COFACTOR.
  4. "Crystal structures of mutant Pseudomonas aeruginosa p-hydroxybenzoate hydroxylases: the Tyr201Phe, Tyr385Phe, and Asn300Asp variants."
    Lah M.S., Palfey B.A., Schreuder H.A., Ludwig M.L.
    Biochemistry 33:1555-1564(1994) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF MUTANTS PHE-201; ASP-300 AND PHE-385 IN COMPLEX WITH SUBSTRATE AND FAD, FUNCTION, MUTAGENESIS OF TYR-201; ASN-300 AND TYR-385, COFACTOR, SUBUNIT.
  5. Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND FAD, FUNCTION, COFACTOR, SUBUNIT.
  6. "pH-dependent structural changes in the active site of p-hydroxybenzoate hydroxylase point to the importance of proton and water movements during catalysis."
    Gatti D.L., Entsch B., Ballou D.P., Ludwig M.L.
    Biochemistry 35:567-578(1996) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG AND FAD, FUNCTION, COFACTOR, SUBUNIT, REACTION MECHANISM.
  7. "Structure-function correlations of the reaction of reduced nicotinamide analogues with p-hydroxybenzoate hydroxylase substituted with a series of 8-substituted flavins."
    Ortiz-Maldonado M., Gatti D., Ballou D.P., Massey V.
    Biochemistry 38:16636-16647(1999) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND FAD ANALOG, FUNCTION, SUBUNIT.
  8. Cited for: X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF MUTANT GLN-220 IN COMPLEX WITH SUBSTRATE AND FAD, FUNCTION, MUTAGENESIS OF ARG-220, COFACTOR, SUBUNIT.
  9. "Removal of a methyl group causes global changes in p-hydroxybenzoate hydroxylase."
    Cole L.J., Gatti D.L., Entsch B., Ballou D.P.
    Biochemistry 44:8047-8058(2005) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF WILD-TYPE AND MUTANT GLY-45 IN COMPLEX WITH SUBSTRATE AND FAD, MUTAGENESIS OF ALA-45, FUNCTION, COFACTOR, SUBUNIT.

Entry informationi

Entry nameiPHHY_PSEAE
AccessioniPrimary (citable) accession number: P20586
Entry historyi
Integrated into UniProtKB/Swiss-Prot: February 1, 1991
Last sequence update: February 1, 1991
Last modified: May 11, 2016
This is version 119 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Miscellaneous

Controlled catalysis is achieved by movement of the flavin and protein between three conformations: in, out and open. The open conformation is important for substrate binding and product release, the in conformation for reaction with oxygen and hydroxylation, and the out conformation for the reduction of FAD by NADPH.1 Publication

Keywords - Technical termi

3D-structure, Complete proteome, Reference proteome

Documents

  1. PATHWAY comments
    Index of metabolic and biosynthesis pathways
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  3. SIMILARITY comments
    Index of protein domains and families

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.