HPAHR_ACIBA
ID HPAHR_ACIBA Reviewed; 315 AA.
AC Q6Q271;
DT 22-FEB-2012, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 25-MAY-2022, entry version 75.
DE RecName: Full=p-hydroxyphenylacetate 3-hydroxylase, reductase component;
DE EC=1.5.1.36 {ECO:0000269|PubMed:15451173};
DE AltName: Full=4-HPA 3-monooxygenase small component {ECO:0000250|UniProtKB:Q5SJP7};
DE AltName: Full=Flavin:NAD(+) oxidoreductase;
DE AltName: Full=p-hydroxyphenylacetate 3-hydroxylase C1 component;
GN Name=C1-hpah {ECO:0000312|EMBL:AAS75430.1};
OS Acinetobacter baumannii.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Moraxellales; Moraxellaceae;
OC Acinetobacter; Acinetobacter calcoaceticus/baumannii complex.
OX NCBI_TaxID=470;
RN [1] {ECO:0000305, ECO:0000312|EMBL:AAS75430.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-27, CATALYTIC
RP ACTIVITY, AND KINETIC PARAMETERS.
RX PubMed=15451173; DOI=10.1016/j.bbaexp.2004.08.003;
RA Thotsaporn K., Sucharitakul J., Wongratana J., Suadee C., Chaiyen P.;
RT "Cloning and expression of p-hydroxyphenylacetate 3-hydroxylase from
RT Acinetobacter baumannii: evidence of the divergence of enzymes in the class
RT of two-protein component aromatic hydroxylases.";
RL Biochim. Biophys. Acta 1680:60-66(2004).
RN [2] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, KINETIC PARAMETERS, SUBUNIT, PATHWAY, AND
RP ACTIVITY REGULATION.
RX PubMed=11683878; DOI=10.1046/j.1432-1033.2001.02490.x;
RA Chaiyen P., Suadee C., Wilairat P.;
RT "A novel two-protein component flavoprotein hydroxylase.";
RL Eur. J. Biochem. 268:5550-5561(2001).
RN [3] {ECO:0000305}
RP FUNCTION, ACTIVITY REGULATION, AND REDOX POTENTIAL.
RX PubMed=16042421; DOI=10.1021/bi050615e;
RA Sucharitakul J., Chaiyen P., Entsch B., Ballou D.P.;
RT "The reductase of p-hydroxyphenylacetate 3-hydroxylase from Acinetobacter
RT baumannii requires p-hydroxyphenylacetate for effective catalysis.";
RL Biochemistry 44:10434-10442(2005).
RN [4] {ECO:0000305}
RP FUNCTION, AND ACTIVITY REGULATION.
RX PubMed=17595116; DOI=10.1021/bi7006614;
RA Sucharitakul J., Phongsak T., Entsch B., Svasti J., Chaiyen P.,
RA Ballou D.P.;
RT "Kinetics of a two-component p-hydroxyphenylacetate hydroxylase explain how
RT reduced flavin is transferred from the reductase to the oxygenase.";
RL Biochemistry 46:8611-8623(2007).
CC -!- FUNCTION: Reductase component of a two-component system that supplies
CC reduced FMN (FMNH2) to the oxygenase component to catalyze the
CC hydroxylation of 4-hydroxyphenylacetic acid, leading to the production
CC of 3,4-dihydroxyphenylacetate (3,4-DHPA). Catalyzes the reduction of
CC free flavins (FMN, FAD and riboflavin) by NADH. Subsequently, the
CC reduced flavins diffuse to the oxygenase component C2.
CC {ECO:0000269|PubMed:11683878, ECO:0000269|PubMed:16042421,
CC ECO:0000269|PubMed:17595116}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a reduced flavin + NAD(+) = an oxidized flavin + 2 H(+) +
CC NADH; Xref=Rhea:RHEA:31303, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945, ChEBI:CHEBI:60531, ChEBI:CHEBI:62787; EC=1.5.1.36;
CC Evidence={ECO:0000269|PubMed:11683878, ECO:0000269|PubMed:15451173};
CC -!- ACTIVITY REGULATION: Flavin concentrations greater than 15 uM do not
CC inhibit the NADH oxidation activity of the reductase component C1 but
CC do affect the hydroxylation activity of the C1-C2 complex. Maximal
CC reductase activity is achieved only upon HPA binding to the reductase
CC component C1 before interaction with NADH. HPA stimulates the rates of
CC both the reduction of FMN and release of reduced FMN from the reductase
CC component. {ECO:0000269|PubMed:11683878, ECO:0000269|PubMed:16042421,
CC ECO:0000269|PubMed:17595116}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=12 uM for NADH (with FMN as cosubstrate)
CC {ECO:0000269|PubMed:11683878};
CC KM=28 uM for NADH (with FAD as cosubstrate)
CC {ECO:0000269|PubMed:11683878};
CC KM=21 uM for NADH (with FMN as cosubstrate)
CC {ECO:0000269|PubMed:15451173};
CC KM=22 uM for NADH (with FAD as cosubstrate)
CC {ECO:0000269|PubMed:15451173};
CC Note=Values measured using the C1-C2 complex. kcat is 343 min(-1) for
CC the reduction of FMN with NADH. {ECO:0000269|PubMed:15451173};
CC Redox potential:
CC E(0) is -236 mV for C1, and -245 mV for C1-HPA complex.
CC {ECO:0000269|PubMed:16042421};
CC -!- PATHWAY: Aromatic compound metabolism; 4-hydroxyphenylacetate
CC degradation; pyruvate and succinate semialdehyde from 4-
CC hydroxyphenylacetate: step 1/7. {ECO:0000269|PubMed:11683878}.
CC -!- SUBUNIT: Homodimer. The p-hydroxyphenylacetate 3-hydroxylase (HpaH) is
CC composed of an oxygenase component C2 and a reductase component C1.
CC {ECO:0000269|PubMed:11683878}.
CC -!- SIMILARITY: Belongs to the non-flavoprotein flavin reductase family.
CC {ECO:0000305}.
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DR EMBL; AY566613; AAS75430.1; -; Genomic_DNA.
DR RefSeq; WP_002047311.1; NZ_VHGP01000058.1.
DR PDB; 5ZC2; X-ray; 2.90 A; A/B=1-315.
DR PDB; 5ZYR; X-ray; 2.20 A; A/B=1-315.
DR PDBsum; 5ZC2; -.
DR PDBsum; 5ZYR; -.
DR AlphaFoldDB; Q6Q271; -.
DR SASBDB; Q6Q271; -.
DR SMR; Q6Q271; -.
DR STRING; 470.IX87_20435; -.
DR PATRIC; fig|470.1289.peg.1766; -.
DR eggNOG; COG1853; Bacteria.
DR BRENDA; 1.14.14.9; 98.
DR UniPathway; UPA00208; UER00416.
DR GO; GO:0036382; F:flavin reductase (NADH) activity; IEA:UniProtKB-EC.
DR GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.10.10.10; -; 1.
DR Gene3D; 2.30.110.10; -; 1.
DR InterPro; IPR002563; Flavin_Rdtase-like_dom.
DR InterPro; IPR012349; Split_barrel_FMN-bd.
DR InterPro; IPR036388; WH-like_DNA-bd_sf.
DR InterPro; IPR036390; WH_DNA-bd_sf.
DR Pfam; PF01613; Flavin_Reduct; 1.
DR SMART; SM00903; Flavin_Reduct; 1.
DR SUPFAM; SSF46785; SSF46785; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aromatic hydrocarbons catabolism; Direct protein sequencing;
KW FAD; Flavoprotein; FMN; NAD; Oxidoreductase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000305|PubMed:15451173"
FT CHAIN 2..315
FT /note="p-hydroxyphenylacetate 3-hydroxylase, reductase
FT component"
FT /id="PRO_0000415754"
FT HELIX 16..23
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 31..36
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 42..47
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 51..54
FT /evidence="ECO:0007829|PDB:5ZYR"
FT TURN 55..58
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 59..65
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 71..76
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 78..84
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 87..89
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 90..95
FT /evidence="ECO:0007829|PDB:5ZYR"
FT TURN 101..106
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 109..111
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 113..115
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 117..121
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 123..136
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 139..151
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 158..160
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 163..168
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 184..186
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 187..189
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 192..213
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 218..227
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 235..242
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 246..258
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 261..264
FT /evidence="ECO:0007829|PDB:5ZYR"
FT STRAND 266..271
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 273..293
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 294..296
FT /evidence="ECO:0007829|PDB:5ZYR"
FT HELIX 299..313
FT /evidence="ECO:0007829|PDB:5ZYR"
SQ SEQUENCE 315 AA; 35412 MW; 33ED493B8B5B0386 CRC64;
MNQLNTAIVE KEVIDPMAFR RALGNFATGV TIMTAQTSSG ERVGVTANSF NSVSLDPALV
LWSIDKKSSS YRIFEEATHF GVNILSAAQI ELSNRFARRS EDKFANIEFD LGVGNIPLFK
NCSAAFECER YNIVEGGDHW IIIGRVVKFH DHGRSPLLYH QGAYSAVLPH PSLNMKSETA
EGVFPGRLYD NMYYLLTQAV RAYQNDYQPK QLASGFRTSE ARLLLVLESK TASSKCDLQR
EVAMPIREIE EATKILSEKG LLIDNGQHYE LTEQGNACAH MLYKIAESHQ EEVFAKYTVD
ERKLFKNMLK DLIGI
