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Entry: PHHY_PSEAE
LinkDB: PHHY_PSEAE
Original site: PHHY_PSEAE 
ID   PHHY_PSEAE              Reviewed;         394 AA.
AC   P20586;
DT   01-FEB-1991, integrated into UniProtKB/Swiss-Prot.
DT   01-FEB-1991, sequence version 1.
DT   16-MAR-2016, entry version 118.
DE   RecName: Full=p-hydroxybenzoate hydroxylase {ECO:0000303|PubMed:1910043};
DE            Short=PHBH {ECO:0000303|PubMed:1910043};
DE            EC=1.14.13.2 {ECO:0000269|PubMed:1910043};
DE   AltName: Full=4-hydroxybenzoate 3-monooxygenase {ECO:0000305};
GN   Name=pobA {ECO:0000303|PubMed:2465205}; OrderedLocusNames=PA0247;
OS   Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG
OS   12228).
OC   Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC   Pseudomonadaceae; Pseudomonas.
OX   NCBI_TaxID=208964;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX   PubMed=2465205; DOI=10.1016/0378-1119(88)90044-3;
RA   Entsch B., Nan Y., Weaich K., Scott K.F.;
RT   "Sequence and organization of pobA, the gene coding for p-
RT   hydroxybenzoate hydroxylase, an inducible enzyme from Pseudomonas
RT   aeruginosa.";
RL   Gene 71:279-291(1988).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228;
RX   PubMed=10984043; DOI=10.1038/35023079;
RA   Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P.,
RA   Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M.,
RA   Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y.,
RA   Brody L.L., Coulter S.N., Folger K.R., Kas A., Larbig K., Lim R.M.,
RA   Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T.,
RA   Reizer J., Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.;
RT   "Complete genome sequence of Pseudomonas aeruginosa PAO1, an
RT   opportunistic pathogen.";
RL   Nature 406:959-964(2000).
RN   [3]
RP   FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF TYR-201 AND TYR-385, AND
RP   COFACTOR.
RX   PubMed=1910043;
RA   Entsch B., Palfey B.A., Ballou D.P., Massey V.;
RT   "Catalytic function of tyrosine residues in para-hydroxybenzoate
RT   hydroxylase as determined by the study of site-directed mutants.";
RL   J. Biol. Chem. 266:17341-17349(1991).
RN   [4]
RP   X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF MUTANTS PHE-201; ASP-300 AND
RP   PHE-385 IN COMPLEX WITH SUBSTRATE AND FAD, FUNCTION, MUTAGENESIS OF
RP   TYR-201; ASN-300 AND TYR-385, COFACTOR, AND SUBUNIT.
RX   PubMed=8312276; DOI=10.1021/bi00172a036;
RA   Lah M.S., Palfey B.A., Schreuder H.A., Ludwig M.L.;
RT   "Crystal structures of mutant Pseudomonas aeruginosa p-hydroxybenzoate
RT   hydroxylases: the Tyr201Phe, Tyr385Phe, and Asn300Asp variants.";
RL   Biochemistry 33:1555-1564(1994).
RN   [5]
RP   X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND
RP   FAD, FUNCTION, COFACTOR, AND SUBUNIT.
RX   PubMed=7939628; DOI=10.1126/science.7939628;
RA   Gatti D.L., Palfey B.A., Lah M.S., Entsch B., Massey V., Ballou D.P.,
RA   Ludwig M.L.;
RT   "The mobile flavin of 4-OH benzoate hydroxylase.";
RL   Science 266:110-114(1994).
RN   [6]
RP   X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEX WITH SUBSTRATE
RP   ANALOG AND FAD, FUNCTION, COFACTOR, SUBUNIT, AND REACTION MECHANISM.
RX   PubMed=8555229; DOI=10.1021/bi951344i;
RA   Gatti D.L., Entsch B., Ballou D.P., Ludwig M.L.;
RT   "pH-dependent structural changes in the active site of p-
RT   hydroxybenzoate hydroxylase point to the importance of proton and
RT   water movements during catalysis.";
RL   Biochemistry 35:567-578(1996).
RN   [7]
RP   X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND
RP   FAD ANALOG, FUNCTION, AND SUBUNIT.
RX   PubMed=10600126; DOI=10.1021/bi991603u;
RA   Ortiz-Maldonado M., Gatti D., Ballou D.P., Massey V.;
RT   "Structure-function correlations of the reaction of reduced
RT   nicotinamide analogues with p-hydroxybenzoate hydroxylase substituted
RT   with a series of 8-substituted flavins.";
RL   Biochemistry 38:16636-16647(1999).
RN   [8]
RP   X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF MUTANT GLN-220 IN COMPLEX
RP   WITH SUBSTRATE AND FAD, FUNCTION, MUTAGENESIS OF ARG-220, COFACTOR,
RP   AND SUBUNIT.
RX   PubMed=11805318; DOI=10.1073/pnas.022640199;
RA   Wang J., Ortiz-Maldonado M., Entsch B., Massey V., Ballou D.,
RA   Gatti D.L.;
RT   "Protein and ligand dynamics in 4-hydroxybenzoate hydroxylase.";
RL   Proc. Natl. Acad. Sci. U.S.A. 99:608-613(2002).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF WILD-TYPE AND MUTANT GLY-45
RP   IN COMPLEX WITH SUBSTRATE AND FAD, MUTAGENESIS OF ALA-45, FUNCTION,
RP   COFACTOR, AND SUBUNIT.
RX   PubMed=15924424; DOI=10.1021/bi050108x;
RA   Cole L.J., Gatti D.L., Entsch B., Ballou D.P.;
RT   "Removal of a methyl group causes global changes in p-hydroxybenzoate
RT   hydroxylase.";
RL   Biochemistry 44:8047-8058(2005).
CC   -!- FUNCTION: Catalyzes the incorporation of an atom of dioxygen into
CC       p-hydroxybenzoate (p-OHB) to form 3,4-dihydroxybenzoate (3,4DOHB).
CC       The reaction occurs in two parts: reduction of the flavin adenine
CC       dinucleotide (FAD) in the enzyme by reduced nicotinamide adenine
CC       dinucleotide phosphate (NADPH) in response to binding p-
CC       hydroxybenzoate to the enzyme and oxidation of reduced FAD with
CC       oxygen to form a hydroperoxide, which then oxygenates p-
CC       hydroxybenzoate. {ECO:0000269|PubMed:10600126,
CC       ECO:0000269|PubMed:11805318, ECO:0000269|PubMed:15924424,
CC       ECO:0000269|PubMed:1910043, ECO:0000269|PubMed:7939628,
CC       ECO:0000269|PubMed:8312276, ECO:0000269|PubMed:8555229}.
CC   -!- CATALYTIC ACTIVITY: 4-hydroxybenzoate + NADPH + O(2) =
CC       protocatechuate + NADP(+) + H(2)O. {ECO:0000269|PubMed:1910043}.
CC   -!- COFACTOR:
CC       Name=FAD; Xref=ChEBI:CHEBI:57692;
CC         Evidence={ECO:0000269|PubMed:11805318,
CC         ECO:0000269|PubMed:15924424, ECO:0000269|PubMed:1910043,
CC         ECO:0000269|PubMed:7939628, ECO:0000269|PubMed:8312276,
CC         ECO:0000269|PubMed:8555229};
CC       Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:11805318,
CC       ECO:0000269|PubMed:15924424, ECO:0000269|PubMed:7939628,
CC       ECO:0000269|PubMed:8312276, ECO:0000269|PubMed:8555229};
CC   -!- PATHWAY: Aromatic compound metabolism; benzoate degradation via
CC       hydroxylation; 3,4-dihydroxybenzoate from benzoate: step 2/2.
CC       {ECO:0000305}.
CC   -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:10600126,
CC       ECO:0000269|PubMed:11805318, ECO:0000269|PubMed:15924424,
CC       ECO:0000269|PubMed:7939628, ECO:0000269|PubMed:8312276,
CC       ECO:0000269|PubMed:8555229}.
CC   -!- MISCELLANEOUS: Controlled catalysis is achieved by movement of the
CC       flavin and protein between three conformations: in, out and open.
CC       The open conformation is important for substrate binding and
CC       product release, the in conformation for reaction with oxygen and
CC       hydroxylation, and the out conformation for the reduction of FAD
CC       by NADPH. {ECO:0000269|PubMed:8555229}.
CC   -!- SIMILARITY: Belongs to the aromatic-ring hydroxylase family.
CC       {ECO:0000305}.
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DR   EMBL; M23173; AAA88455.1; -; Genomic_DNA.
DR   EMBL; AE004091; AAG03636.1; -; Genomic_DNA.
DR   PIR; JT0384; WHPSBA.
DR   RefSeq; NP_248938.1; NC_002516.2.
DR   RefSeq; WP_003112685.1; NZ_ASJY01000056.1.
DR   PDB; 1D7L; X-ray; 2.20 A; A=1-394.
DR   PDB; 1DOB; X-ray; 2.00 A; A=1-394.
DR   PDB; 1DOC; X-ray; 2.00 A; A=1-394.
DR   PDB; 1DOD; X-ray; 2.10 A; A=1-394.
DR   PDB; 1DOE; X-ray; 2.30 A; A=1-394.
DR   PDB; 1IUS; X-ray; 2.20 A; A=1-394.
DR   PDB; 1IUT; X-ray; 2.00 A; A=1-394.
DR   PDB; 1IUU; X-ray; 2.00 A; A=1-394.
DR   PDB; 1IUV; X-ray; 2.50 A; A=1-394.
DR   PDB; 1IUW; X-ray; 2.00 A; A=1-394.
DR   PDB; 1IUX; X-ray; 2.00 A; A=1-394.
DR   PDB; 1K0I; X-ray; 1.80 A; A=1-394.
DR   PDB; 1K0J; X-ray; 2.20 A; A=1-394.
DR   PDB; 1K0L; X-ray; 2.00 A; A=1-394.
DR   PDB; 1PXA; X-ray; 2.30 A; A=1-394.
DR   PDB; 1PXB; X-ray; 2.30 A; A=1-394.
DR   PDB; 1PXC; X-ray; 2.10 A; A=1-394.
DR   PDB; 1YKJ; X-ray; 2.00 A; A/B=1-394.
DR   PDBsum; 1D7L; -.
DR   PDBsum; 1DOB; -.
DR   PDBsum; 1DOC; -.
DR   PDBsum; 1DOD; -.
DR   PDBsum; 1DOE; -.
DR   PDBsum; 1IUS; -.
DR   PDBsum; 1IUT; -.
DR   PDBsum; 1IUU; -.
DR   PDBsum; 1IUV; -.
DR   PDBsum; 1IUW; -.
DR   PDBsum; 1IUX; -.
DR   PDBsum; 1K0I; -.
DR   PDBsum; 1K0J; -.
DR   PDBsum; 1K0L; -.
DR   PDBsum; 1PXA; -.
DR   PDBsum; 1PXB; -.
DR   PDBsum; 1PXC; -.
DR   PDBsum; 1YKJ; -.
DR   ProteinModelPortal; P20586; -.
DR   SMR; P20586; 1-394.
DR   STRING; 208964.PA0247; -.
DR   DrugBank; DB03147; Flavin adenine dinucleotide.
DR   PaxDb; P20586; -.
DR   EnsemblBacteria; AAG03636; AAG03636; PA0247.
DR   GeneID; 882128; -.
DR   KEGG; pae:PA0247; -.
DR   PATRIC; 19834710; VBIPseAer58763_0257.
DR   PseudoCAP; PA0247; -.
DR   eggNOG; ENOG41072AC; Bacteria.
DR   eggNOG; COG0654; LUCA.
DR   HOGENOM; HOG000221233; -.
DR   InParanoid; P20586; -.
DR   KO; K00481; -.
DR   OMA; VEPMQYG; -.
DR   OrthoDB; EOG6PS5RW; -.
DR   PhylomeDB; P20586; -.
DR   BRENDA; 1.14.13.2; 5087.
DR   UniPathway; UPA00156; UER00257.
DR   EvolutionaryTrace; P20586; -.
DR   Proteomes; UP000002438; Chromosome.
DR   GO; GO:0018659; F:4-hydroxybenzoate 3-monooxygenase activity; IEA:UniProtKB-EC.
DR   GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR   GO; GO:0004497; F:monooxygenase activity; IBA:GO_Central.
DR   GO; GO:0043640; P:benzoate catabolic process via hydroxylation; IEA:UniProtKB-UniPathway.
DR   GO; GO:0044550; P:secondary metabolite biosynthetic process; IBA:GO_Central.
DR   Gene3D; 3.50.50.60; -; 1.
DR   InterPro; IPR002938; FAD-bd.
DR   InterPro; IPR023753; FAD/NAD-binding_dom.
DR   InterPro; IPR012733; HB_mOase.
DR   Pfam; PF01494; FAD_binding_3; 1.
DR   SUPFAM; SSF51905; SSF51905; 2.
DR   TIGRFAMs; TIGR02360; pbenz_hydroxyl; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Aromatic hydrocarbons catabolism; Complete proteome;
KW   FAD; Flavoprotein; Monooxygenase; NADP; Oxidoreductase;
KW   Reference proteome.
FT   CHAIN         1    394       p-hydroxybenzoate hydroxylase.
FT                                /FTId=PRO_0000058381.
FT   NP_BIND      42     47       FAD. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   NP_BIND     299    300       FAD. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   REGION      212    214       Substrate binding.
FT                                {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING      13     13       FAD. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING      32     32       FAD. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING     102    102       FAD. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING     201    201       Substrate. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING     222    222       Substrate. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING     286    286       FAD. {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   BINDING     293    293       Substrate; via carbonyl oxygen.
FT                                {ECO:0000269|PubMed:10600126,
FT                                ECO:0000269|PubMed:11805318,
FT                                ECO:0000269|PubMed:15924424,
FT                                ECO:0000269|PubMed:7939628,
FT                                ECO:0000269|PubMed:8312276,
FT                                ECO:0000269|PubMed:8555229}.
FT   SITE        201    201       Important for catalytic activity.
FT                                {ECO:0000269|PubMed:8312276}.
FT   SITE        385    385       Important for catalytic activity.
FT                                {ECO:0000269|PubMed:8312276}.
FT   MUTAGEN      45     45       A->G: The positions of the substrate and
FT                                the flavin are not altered.
FT                                {ECO:0000269|PubMed:15924424}.
FT   MUTAGEN     201    201       Y->F: Reduction of hydroxylase activity.
FT                                {ECO:0000269|PubMed:1910043,
FT                                ECO:0000269|PubMed:8312276}.
FT   MUTAGEN     220    220       R->Q: Lower affinity for p-OHB than the
FT                                wild-type. {ECO:0000269|PubMed:11805318}.
FT   MUTAGEN     300    300       N->D: The side chain of Asp300 moves away
FT                                from the flavin, disrupting the
FT                                interactions of the carboxamide group
FT                                with the flavin O(2) atom, and the alpha-
FT                                helix H10 that begins at residue 297 is
FT                                displaced, altering its dipole
FT                                interactions with the flavin ring.
FT                                {ECO:0000269|PubMed:8312276}.
FT   MUTAGEN     385    385       Y->F: The positions of the substrate and
FT                                the flavin are not altered.
FT                                {ECO:0000269|PubMed:1910043,
FT                                ECO:0000269|PubMed:8312276}.
FT   STRAND        4      8       {ECO:0000244|PDB:1K0I}.
FT   HELIX        12     24       {ECO:0000244|PDB:1K0I}.
FT   STRAND       28     31       {ECO:0000244|PDB:1K0I}.
FT   HELIX        36     40       {ECO:0000244|PDB:1K0I}.
FT   STRAND       47     49       {ECO:0000244|PDB:1DOB}.
FT   HELIX        50     58       {ECO:0000244|PDB:1K0I}.
FT   HELIX        63     68       {ECO:0000244|PDB:1K0I}.
FT   STRAND       70     73       {ECO:0000244|PDB:1K0I}.
FT   STRAND       75     79       {ECO:0000244|PDB:1K0I}.
FT   STRAND       82     86       {ECO:0000244|PDB:1K0I}.
FT   HELIX        88     92       {ECO:0000244|PDB:1K0I}.
FT   STRAND       97     99       {ECO:0000244|PDB:1K0I}.
FT   HELIX       102    115       {ECO:0000244|PDB:1K0I}.
FT   STRAND      119    123       {ECO:0000244|PDB:1K0I}.
FT   STRAND      125    130       {ECO:0000244|PDB:1K0I}.
FT   STRAND      134    136       {ECO:0000244|PDB:1K0I}.
FT   STRAND      138    143       {ECO:0000244|PDB:1K0I}.
FT   STRAND      146    151       {ECO:0000244|PDB:1K0I}.
FT   STRAND      153    157       {ECO:0000244|PDB:1K0I}.
FT   HELIX       166    168       {ECO:0000244|PDB:1K0I}.
FT   HELIX       171    173       {ECO:0000244|PDB:1K0I}.
FT   STRAND      175    192       {ECO:0000244|PDB:1K0I}.
FT   STRAND      195    198       {ECO:0000244|PDB:1DOB}.
FT   STRAND      200    202       {ECO:0000244|PDB:1K0I}.
FT   STRAND      209    215       {ECO:0000244|PDB:1K0I}.
FT   STRAND      218    225       {ECO:0000244|PDB:1K0I}.
FT   HELIX       231    233       {ECO:0000244|PDB:1K0I}.
FT   HELIX       236    245       {ECO:0000244|PDB:1K0I}.
FT   HELIX       249    254       {ECO:0000244|PDB:1K0I}.
FT   STRAND      260    274       {ECO:0000244|PDB:1K0I}.
FT   STRAND      276    278       {ECO:0000244|PDB:1K0I}.
FT   STRAND      281    283       {ECO:0000244|PDB:1K0I}.
FT   HELIX       285    287       {ECO:0000244|PDB:1K0I}.
FT   HELIX       293    295       {ECO:0000244|PDB:1K0I}.
FT   HELIX       298    319       {ECO:0000244|PDB:1K0I}.
FT   HELIX       322    327       {ECO:0000244|PDB:1K0I}.
FT   HELIX       328    350       {ECO:0000244|PDB:1K0I}.
FT   HELIX       358    373       {ECO:0000244|PDB:1K0I}.
FT   HELIX       375    385       {ECO:0000244|PDB:1K0I}.
SQ   SEQUENCE   394 AA;  44324 MW;  1E7232854D9EC792 CRC64;
     MKTQVAIIGA GPSGLLLGQL LHKAGIDNVI LERQTPDYVL GRIRAGVLEQ GMVDLLREAG
     VDRRMARDGL VHEGVEIAFA GQRRRIDLKR LSGGKTVTVY GQTEVTRDLM EAREACGATT
     VYQAAEVRLH DLQGERPYVT FERDGERLRL DCDYIAGCDG FHGISRQSIP AERLKVFERV
     YPFGWLGLLA DTPPVSHELI YANHPRGFAL CSQRSATRSR YYVQVPLSEK VEDWSDERFW
     TELKARLPSE VAEKLVTGPS LEKSIAPLRS FVVEPMQHGR LFLAGDAAHI VPPTGAKGLN
     LAASDVSTLY RLLLKAYREG RGELLERYSA ICLRRIWKAE RFSWWMTSVL HRFPDTDAFS
     QRIQQTELEY YLGSEAGLAT IAENYVGLPY EEIE
//
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