PAO_PSESP
ID PAO_PSESP Reviewed; 714 AA.
AC Q5W9R9; Q7M1A6;
DT 16-OCT-2013, integrated into UniProtKB/Swiss-Prot.
DT 07-DEC-2004, sequence version 1.
DT 03-AUG-2022, entry version 68.
DE RecName: Full=Phenylalanine 2-monooxygenase precursor;
DE Short=proPAO;
DE EC=1.13.12.9;
DE AltName: Full=L-phenylalanine oxidase (deaminating and decarboxylating);
DE Short=PAO;
DE Contains:
DE RecName: Full=Phenylalanine 2-monooxygenase alpha subunit;
DE Contains:
DE RecName: Full=Phenylalanine 2-monooxygenase beta subunit;
DE Flags: Precursor;
OS Pseudomonas sp.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=306;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, COFACTOR, AND PROTEOLYTIC
RP PROCESSING.
RC STRAIN=P-501;
RX PubMed=15632301; DOI=10.1093/jb/mvh169;
RA Suzuki H., Higashi Y., Asano M., Suguro M., Kigawa M., Maeda M.,
RA Katayama S., Mukouyama E.B., Uchiyama K.;
RT "Sequencing and expression of the L-phenylalanine oxidase gene from
RT Pseudomonas sp. P-501. Proteolytic activation of the proenzyme.";
RL J. Biochem. 136:617-627(2004).
RN [2]
RP PROTEIN SEQUENCE OF 16-107, AND SUBUNIT.
RC STRAIN=P-501;
RX PubMed=8109968; DOI=10.1006/abbi.1994.1056;
RA Mukouyama E.B., Suzuki H., Koyama H.;
RT "New subunit in L-phenylalanine oxidase from Pseudomonas sp. P-501 and the
RT primary structure.";
RL Arch. Biochem. Biophys. 308:400-406(1994).
RN [3]
RP PROTEIN SEQUENCE OF 110-183.
RX PubMed=9603998; DOI=10.1093/oxfordjournals.jbchem.a022048;
RA Mukouyama E.B., Hirose T., Suzuki H.;
RT "Chemical modification of L-phenylalanine oxidase from Pseudomonas sp. P-
RT 501 by phenylglyoxal. Identification of one essential arginyl residue.";
RL J. Biochem. 123:1097-1103(1998).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND SUBSTRATE SPECIFICITY.
RX PubMed=7174643; DOI=10.1093/oxfordjournals.jbchem.a134041;
RA Koyama H.;
RT "Purification and characterization of a novel L-phenylalanine oxidase
RT (deaminating and decarboxylating) from Pseudomonas sp. P-501.";
RL J. Biochem. 92:1235-1240(1982).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=6885723; DOI=10.1093/oxfordjournals.jbchem.a134265;
RA Koyama H.;
RT "Further characterization of a novel L-phenylalanine oxidase (deaminating
RT and decarboxylating) from Pseudomonas sp. P-501.";
RL J. Biochem. 93:1313-1319(1983).
RN [6]
RP FUNCTION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=6501250; DOI=10.1093/oxfordjournals.jbchem.a134853;
RA Koyama H.;
RT "Oxidation and oxygenation of L-amino acids catalyzed by a L-phenylalanine
RT oxidase (deaminating and decarboxylating) from Pseudomonas sp. P-501.";
RL J. Biochem. 96:421-427(1984).
RN [7]
RP FUNCTION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=3818566; DOI=10.1093/oxfordjournals.jbchem.a121798;
RA Koyama H., Suzuki H.;
RT "Spectral and kinetic studies on Pseudomonas L-phenylalanine oxidase
RT (deaminating and decarboxylating).";
RL J. Biochem. 100:859-866(1986).
RN [8]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=16567420; DOI=10.1093/jb/mvj049;
RA Ohta Y., Mukouyama E.B., Suzuki H.;
RT "Kinetic isotope effect of the L-phenylalanine oxidase from Pseudomonas sp.
RT P-501.";
RL J. Biochem. 139:551-555(2006).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (1.25 ANGSTROMS) OF 2-714 IN COMPLEX WITH FAD AND
RP SUBSTRATE ANALOG, AND MUTAGENESIS OF MET-143 AND LYS-479.
RC STRAIN=P-501;
RX PubMed=18417467; DOI=10.1074/jbc.m800366200;
RA Ida K., Kurabayashi M., Suguro M., Hiruma Y., Hikima T., Yamomoto M.,
RA Suzuki H.;
RT "Structural basis of proteolytic activation of L-phenylalanine oxidase from
RT Pseudomonas sp. P-501.";
RL J. Biol. Chem. 283:16584-16590(2008).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (1.10 ANGSTROMS) OF 2-714 IN COMPLEXES WITH
RP L-PHENYLALANINE AND L-METHIONINE, AND MUTAGENESIS OF ARG-144 AND TYR-537.
RC STRAIN=P-501;
RX PubMed=21841183; DOI=10.1093/jb/mvr103;
RA Ida K., Suguro M., Suzuki H.;
RT "High resolution X-ray crystal structures of L-phenylalanine oxidase
RT (deaminating and decarboxylating) from Pseudomonas sp. P-501. Structures of
RT the enzyme-ligand complex and catalytic mechanism.";
RL J. Biochem. 150:659-669(2011).
CC -!- FUNCTION: Catalyzes both oxygenative decarboxylation and oxidative
CC deamination, depending on the substrate used. Has high activity for L-
CC Phe and L-Tyr, but relatively low activities for L-Met and L-Trp. L-Phe
CC is mainly oxygenated and L-Met is mainly oxidized.
CC {ECO:0000269|PubMed:15632301, ECO:0000269|PubMed:16567420,
CC ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723, ECO:0000269|PubMed:7174643}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-phenylalanine + O2 = 2-phenylacetamide + CO2 + H2O;
CC Xref=Rhea:RHEA:10712, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:16562, ChEBI:CHEBI:58095;
CC EC=1.13.12.9; Evidence={ECO:0000269|PubMed:16567420,
CC ECO:0000269|PubMed:6885723, ECO:0000269|PubMed:7174643};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:15632301, ECO:0000269|PubMed:6885723};
CC Note=Binds 2 FAD per tetramer. {ECO:0000269|PubMed:15632301,
CC ECO:0000269|PubMed:6885723};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=11.1 uM for L-Phe (for the oxygenation reaction)
CC {ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723};
CC KM=13.3 uM for L-Phe (for the oxidation reaction)
CC {ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723};
CC KM=4 mM for L-Tyr {ECO:0000269|PubMed:3818566,
CC ECO:0000269|PubMed:6501250, ECO:0000269|PubMed:6885723};
CC KM=2.2 mM for L-Met {ECO:0000269|PubMed:3818566,
CC ECO:0000269|PubMed:6501250, ECO:0000269|PubMed:6885723};
CC KM=1.96 mM for O(2) (for the oxygenation reaction with L-Phe as
CC cosubstrate) {ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723};
CC KM=2.04 mM for O(2) (for the oxidation reaction with L-Phe as
CC cosubstrate) {ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723};
CC KM=3.145 mM for O(2) (with L-Tyr as cosubstrate)
CC {ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723};
CC KM=1.258 mM for O(2) (with L-Met as cosubstrate)
CC {ECO:0000269|PubMed:3818566, ECO:0000269|PubMed:6501250,
CC ECO:0000269|PubMed:6885723};
CC pH dependence:
CC Optimum pH is 6-9 for the oxygenation reaction and 10.5 for the
CC oxidation reaction. {ECO:0000269|PubMed:3818566,
CC ECO:0000269|PubMed:6501250, ECO:0000269|PubMed:6885723};
CC Temperature dependence:
CC Optimum temperature is 45 degrees Celsius for the oxygenation
CC reaction and 65 degrees Celsius for the oxygenation reaction. Stable
CC up to 70 degrees Celsius. {ECO:0000269|PubMed:3818566,
CC ECO:0000269|PubMed:6501250, ECO:0000269|PubMed:6885723};
CC -!- SUBUNIT: Heterotetramer composed of 2 alpha and 2 beta subunits.
CC {ECO:0000269|PubMed:18417467, ECO:0000269|PubMed:8109968}.
CC -!- PTM: Proteolytically cleaved to yield the active enzyme. Cleavage of
CC the linkage between the 2 subunits causes reshaping of the oxygen
CC channel and the hydrophobic environment around the flavin ring. Removal
CC of the prosequence causes opening of the amino acid channel.
CC {ECO:0000269|PubMed:15632301}.
CC -!- SIMILARITY: Belongs to the phenylalanine 2-monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; AB167410; BAD66877.1; -; Genomic_DNA.
DR PIR; S41662; S41662.
DR PDB; 2YR4; X-ray; 1.70 A; A/B=2-714.
DR PDB; 2YR5; X-ray; 1.25 A; A/B=2-714.
DR PDB; 2YR6; X-ray; 1.35 A; A/B=2-714.
DR PDB; 3AYI; X-ray; 1.25 A; A/B=2-714.
DR PDB; 3AYJ; X-ray; 1.10 A; A/B=2-714.
DR PDB; 3AYL; X-ray; 1.25 A; A/B=2-714.
DR PDBsum; 2YR4; -.
DR PDBsum; 2YR5; -.
DR PDBsum; 2YR6; -.
DR PDBsum; 3AYI; -.
DR PDBsum; 3AYJ; -.
DR PDBsum; 3AYL; -.
DR AlphaFoldDB; Q5W9R9; -.
DR SMR; Q5W9R9; -.
DR KEGG; ag:BAD66877; -.
DR BRENDA; 1.13.12.9; 5085.
DR EvolutionaryTrace; Q5W9R9; -.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0050172; F:phenylalanine 2-monooxygenase activity; IDA:UniProtKB.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; FAD; Flavoprotein; Monooxygenase;
KW Nucleotide-binding; Oxidoreductase; Zymogen.
FT PROPEP 1..15
FT /note="Removed in mature form; occupies the channel of the
FT substrate amino acid from the outside of the protein to the
FT interior flavin ring in the precursor"
FT /evidence="ECO:0000269|PubMed:8109968"
FT /id="PRO_0000424222"
FT CHAIN 16..107
FT /note="Phenylalanine 2-monooxygenase alpha subunit"
FT /id="PRO_5000051132"
FT PROPEP 108..109
FT /note="Linker peptide"
FT /evidence="ECO:0000269|PubMed:9603998"
FT /id="PRO_0000424223"
FT CHAIN 110..714
FT /note="Phenylalanine 2-monooxygenase beta subunit"
FT /id="PRO_0000424224"
FT BINDING 2
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 68
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 95..96
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 120
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 141..144
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 144
FT /ligand="substrate"
FT BINDING 375
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 537
FT /ligand="substrate"
FT BINDING 652..653
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 660..662
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:18417467"
FT BINDING 660
FT /ligand="substrate"
FT MUTAGEN 143
FT /note="M->A: Reduces catalytic efficiency 10-fold."
FT /evidence="ECO:0000269|PubMed:18417467"
FT MUTAGEN 144
FT /note="R->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:21841183"
FT MUTAGEN 144
FT /note="R->K: Reduces catalytic activity 400-fold."
FT /evidence="ECO:0000269|PubMed:21841183"
FT MUTAGEN 479
FT /note="K->A: Reduces catalytic efficiency 200-fold."
FT /evidence="ECO:0000269|PubMed:18417467"
FT MUTAGEN 537
FT /note="Y->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:21841183"
FT MUTAGEN 537
FT /note="Y->F: Reduces catalytic activity 17-fold."
FT /evidence="ECO:0000269|PubMed:21841183"
FT CONFLICT 178
FT /note="E -> W (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT HELIX 22..28
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 29..31
FT /evidence="ECO:0007829|PDB:2YR5"
FT HELIX 35..37
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 42..45
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 55..63
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 67..80
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 88..94
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 101..103
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 121..126
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 131..133
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 135..139
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 150..160
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 177..181
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 184..191
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 193..195
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 196..198
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 202..214
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 218..221
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 233..240
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 247..255
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 257..265
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 270..283
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 290..292
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 296..310
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 312..315
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 318..320
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 325..333
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 334..337
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 339..341
FT /evidence="ECO:0007829|PDB:2YR4"
FT STRAND 343..346
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 348..361
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 362..364
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 365..371
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 373..380
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 382..385
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 388..394
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 399..409
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 413..420
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 421..423
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 428..434
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 436..438
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 443..449
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 459..470
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 477..485
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 486..490
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 504..507
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 508..511
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 512..518
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 529..539
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 541..547
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 552..561
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 565..574
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 591..597
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 605..608
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 609..611
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 615..619
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 623..625
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 626..634
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 635..640
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 642..644
FT /evidence="ECO:0007829|PDB:3AYJ"
FT STRAND 648..650
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 653..655
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 662..680
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 681..683
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 685..687
FT /evidence="ECO:0007829|PDB:3AYJ"
FT TURN 690..693
FT /evidence="ECO:0007829|PDB:3AYJ"
FT HELIX 694..697
FT /evidence="ECO:0007829|PDB:3AYJ"
SQ SEQUENCE 714 AA; 76883 MW; 2EBEA742F1D41189 CRC64;
MGVTVIPRLL GLKDEKKIAT TVGEARLSGI NYRHPDSALV SYPVAAAAPL GRLPAGNYRI
AIVGGGAGGI AALYELGRLA ATLPAGSGID VQIYEADPDS FLHDRPGIKA IKVRGLKAGR
VSAALVHNGD PASGDTIYEV GAMRFPEIAG LTWHYASAAF GDAAPIKVFP NPGKVPTEFV
FGNRVDRYVG SDPKDWEDPD SPTLKVLGVV AGGLVGNPQG ENVAMYPIAN VDPAKIAAIL
NAATPPADAL ERIQTKYWPE FIAQYDGLTL GAAVREIVTV AFEKGTLPPV DGVLDVDESI
SYYVELFGRF GFGTGGFKPL YNISLVEMMR LILWDYSNEY TLPVTENVEF IRNLFLKAQN
VGAGKLVVQV RQERVANACH SGTASARAQL LSYDSHNAVH SEAYDFVILA VPHDQLTPIV
SRSGFEHAAS QNLGDAGLGL ETHTYNQVYP PLLLSDSSPA ANARIVTAIG QLHMARSSKV
FATVKTAALD QPWVPQWRGE PIKAVVSDSG LAASYVVPSP IVEDGQAPEY SSLLASYTWE
DDSTRLRHDF GLYPQNPATE TGTADGMYRT MVNRAYRYVK YAGASNAQPW WFYQLLAEAR
TADRFVFDWT TNKTAGGFKL DMTGDHHQSN LCFRYHTHAL AASLDNRFFI ASDSYSHLGG
WLEGAFMSAL NAVAGLIVRA NRGDVSALST EARPLVIGLR PVVKVPAAEL ATSQ
