PHSA_STRAT
ID PHSA_STRAT Reviewed; 643 AA.
AC Q53692;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 97.
DE RecName: Full=O-aminophenol oxidase {ECO:0000303|PubMed:2477054};
DE EC=1.10.3.4 {ECO:0000269|PubMed:2477054, ECO:0000269|PubMed:4118295};
DE AltName: Full=Phenoxazinone synthase {ECO:0000303|PubMed:4118295};
DE Short=PHS {ECO:0000303|PubMed:4118295};
GN Name=phsA {ECO:0000303|PubMed:7592317};
OS Streptomyces antibioticus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1890;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 2-16.
RC STRAIN=DSM 41481 / IMRU 3720;
RX PubMed=7592317; DOI=10.1128/jb.177.20.5740-5747.1995;
RA Hsieh C.-J., Jones G.H.;
RT "Nucleotide sequence, transcriptional analysis, and glucose regulation of
RT the phenoxazinone synthase gene (phsA) from Streptomyces antibioticus.";
RL J. Bacteriol. 177:5740-5747(1995).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBSTRATE
RP SPECIFICITY.
RX PubMed=4118295; DOI=10.1128/jb.112.3.1353-1357.1972;
RA Golub E.E., Nishimura J.S.;
RT "Phenoxazinone synthetase from Streptomyces antibioticus: multiple
RT activities of the enzyme.";
RL J. Bacteriol. 112:1353-1357(1972).
RN [3]
RP SUBUNIT.
RX PubMed=7305384; DOI=10.1016/0003-9861(81)90429-x;
RA Choy H.A., Jones G.H.;
RT "Phenoxazinone synthase from Streptomyces antibiotics: purification of the
RT large and small enzyme forms.";
RL Arch. Biochem. Biophys. 211:55-65(1981).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP REACTION MECHANISM.
RX PubMed=2477054; DOI=10.1021/bi00441a026;
RA Barry C.E. III, Nayar P.G., Begley T.P.;
RT "Phenoxazinone synthase: mechanism for the formation of the phenoxazinone
RT chromophore of actinomycin.";
RL Biochemistry 28:6323-6333(1989).
RN [5]
RP CRYSTALLIZATION, AND SUBUNIT.
RX PubMed=15272175; DOI=10.1107/s0907444904013204;
RA Smith A.W., Camara-Artigas A., Olea C. Jr., Francisco W.A., Allen J.P.;
RT "Crystallization and initial X-ray analysis of phenoxazinone synthase from
RT Streptomyces antibioticus.";
RL Acta Crystallogr. D 60:1453-1455(2004).
RN [6]
RP FUNCTION.
RX PubMed=10770769; DOI=10.1128/aac.44.5.1322-1327.2000;
RA Jones G.H.;
RT "Actinomycin production persists in a strain of Streptomyces antibioticus
RT lacking phenoxazinone synthase.";
RL Antimicrob. Agents Chemother. 44:1322-1327(2000).
RN [7]
RP REVIEW, FUNCTION, AND COFACTOR.
RX PubMed=19268377; DOI=10.1016/j.tibtech.2009.01.001;
RA Le Roes-Hill M., Goodwin C., Burton S.;
RT "Phenoxazinone synthase: what's in a name?";
RL Trends Biotechnol. 27:248-258(2009).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 32-643 IN COMPLEX WITH COPPER
RP IONS, SUBUNIT, AND COFACTOR.
RX PubMed=16584173; DOI=10.1021/bi0525526;
RA Smith A.W., Camara-Artigas A., Wang M., Allen J.P., Francisco W.A.;
RT "Structure of phenoxazinone synthase from Streptomyces antibioticus reveals
RT a new type 2 copper center.";
RL Biochemistry 45:4378-4387(2006).
CC -!- FUNCTION: Could be involved in the spore pigmentation and melanin
CC production. Catalyzes the oxidative coupling of 2-aminophenols to form
CC the 2-aminophenoxazinone chromophore. 2-aminophenoxazinone synthesis
CC proceeds via a sequence of three consecutive 2-electron aminophenol
CC oxidations. First, the o-aminophenol is oxidized by two electrons to
CC the quinone imine, which then conjugates to a second o-aminophenol
CC molecule while still bound to the enzyme. This product is further
CC oxidized by two electrons to give rise to the p-quinone imine. The last
CC two steps of the reaction, another conjugation to generate the
CC tricyclic structure and a final two-electron oxidation to yield the 2-
CC aminophenoxazinone product, are thought to be non-enzymatic. It can
CC also uuse 3-hydroxyanthranilic acid (HAA), 4-methyl-3-
CC hydroxyanthranilic acid (MHA), 3,4-dihydroxy-L-phenylalanine (L-DOPA),
CC ferrocyanide and thiophenol as substrates.
CC {ECO:0000269|PubMed:10770769, ECO:0000269|PubMed:2477054,
CC ECO:0000269|PubMed:4118295, ECO:0000303|PubMed:19268377}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4 2-aminophenol + 3 O2 = 2 2-aminophenoxazin-3-one + 6 H2O;
CC Xref=Rhea:RHEA:40963, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17293, ChEBI:CHEBI:18112; EC=1.10.3.4;
CC Evidence={ECO:0000269|PubMed:2477054, ECO:0000269|PubMed:4118295};
CC -!- COFACTOR:
CC Name=Cu(2+); Xref=ChEBI:CHEBI:29036;
CC Evidence={ECO:0000269|PubMed:16584173, ECO:0000269|PubMed:2477054,
CC ECO:0000303|PubMed:19268377};
CC Note=Binds 5 copper ions per monomer. {ECO:0000269|PubMed:16584173,
CC ECO:0000269|PubMed:2477054, ECO:0000303|PubMed:19268377};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=207 uM for dioxygen {ECO:0000269|PubMed:2477054};
CC KM=240 uM for MHA {ECO:0000269|PubMed:4118295};
CC KM=400 uM for HAA {ECO:0000269|PubMed:4118295};
CC KM=400 uM for thiophenol {ECO:0000269|PubMed:4118295};
CC KM=500 uM for L-DOPA {ECO:0000269|PubMed:4118295};
CC KM=500 uM for 2-aminophenol {ECO:0000269|PubMed:2477054};
CC KM=710 uM for ferrocyanide {ECO:0000269|PubMed:4118295};
CC Vmax=8.6 umol/min/mg enzyme toward 2-aminophenol
CC {ECO:0000269|PubMed:2477054};
CC Vmax=36.6 umol/min/mg enzyme toward dioxygen
CC {ECO:0000269|PubMed:2477054};
CC pH dependence:
CC Optimum pH is 5. {ECO:0000269|PubMed:2477054};
CC -!- SUBUNIT: Homodimer (small form) or homohexamer (large form).
CC {ECO:0000269|PubMed:15272175, ECO:0000269|PubMed:16584173,
CC ECO:0000269|PubMed:7305384}.
CC -!- SIMILARITY: Belongs to the multicopper oxidase family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAA86668.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
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DR EMBL; U04283; AAA86668.1; ALT_INIT; Unassigned_DNA.
DR PDB; 3GYR; X-ray; 2.30 A; A/B/C/D/E/F/G/H/I/J/K/L=32-643.
DR PDBsum; 3GYR; -.
DR AlphaFoldDB; Q53692; -.
DR SMR; Q53692; -.
DR STRING; 1890.AFM16_29460; -.
DR KEGG; ag:AAA86668; -.
DR BioCyc; MetaCyc:MON-19511; -.
DR BRENDA; 1.10.3.4; 5974.
DR EvolutionaryTrace; Q53692; -.
DR GO; GO:0005507; F:copper ion binding; IEA:InterPro.
DR GO; GO:0050149; F:o-aminophenol oxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 2.60.40.420; -; 3.
DR InterPro; IPR011706; Cu-oxidase_C.
DR InterPro; IPR045087; Cu-oxidase_fam.
DR InterPro; IPR011707; Cu-oxidase_N.
DR InterPro; IPR033138; Cu_oxidase_CS.
DR InterPro; IPR002355; Cu_oxidase_Cu_BS.
DR InterPro; IPR008972; Cupredoxin.
DR PANTHER; PTHR11709; PTHR11709; 1.
DR Pfam; PF07731; Cu-oxidase_2; 1.
DR Pfam; PF07732; Cu-oxidase_3; 2.
DR SUPFAM; SSF49503; SSF49503; 3.
DR PROSITE; PS00079; MULTICOPPER_OXIDASE1; 1.
DR PROSITE; PS00080; MULTICOPPER_OXIDASE2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Copper; Direct protein sequencing;
KW Metal-binding; Oxidoreductase; Repeat.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:7592317"
FT CHAIN 2..643
FT /note="O-aminophenol oxidase"
FT /id="PRO_0000085583"
FT DOMAIN 87..224
FT /note="Plastocyanin-like 1"
FT /evidence="ECO:0000255"
FT DOMAIN 494..621
FT /note="Plastocyanin-like 2"
FT /evidence="ECO:0000255"
FT REGION 129..149
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 162
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="1"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 164
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="2"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 202
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="2"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 204
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="3"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 435
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 439
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 441
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 525
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="5"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 528
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="1"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 530
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="3"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 603
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="3"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 604
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="5"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 605
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="2"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 609
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="5"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT BINDING 614
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="5"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:16584173"
FT STRAND 66..75
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 84..89
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 92..94
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 97..101
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 106..112
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 121..127
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 137..139
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 150..153
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 161..164
FT /evidence="ECO:0007829|PDB:3GYR"
FT TURN 170..172
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 176..178
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 185..190
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 196..203
FT /evidence="ECO:0007829|PDB:3GYR"
FT TURN 206..208
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 209..212
FT /evidence="ECO:0007829|PDB:3GYR"
FT TURN 213..216
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 218..224
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 226..229
FT /evidence="ECO:0007829|PDB:3GYR"
FT TURN 230..232
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 236..238
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 239..247
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 249..251
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 257..271
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 273..276
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 286..290
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 293..295
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 297..311
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 318..323
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 333..339
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 342..350
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 352..361
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 366..372
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 380..385
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 397..399
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 405..412
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 440..447
FT /evidence="ECO:0007829|PDB:3GYR"
FT TURN 452..456
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 459..465
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 479..483
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 489..496
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 505..508
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 512..519
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 521..523
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 525..531
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 533..543
FT /evidence="ECO:0007829|PDB:3GYR"
FT TURN 549..552
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 553..556
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 558..565
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 576..581
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 583..592
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 598..606
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 607..611
FT /evidence="ECO:0007829|PDB:3GYR"
FT STRAND 615..621
FT /evidence="ECO:0007829|PDB:3GYR"
FT HELIX 623..626
FT /evidence="ECO:0007829|PDB:3GYR"
SQ SEQUENCE 643 AA; 70245 MW; FB6D9208221BDAB8 CRC64;
MTDMIEQSDD RIDPIDGVLA DGVLADDVLA KEREQAPAPG ELTPFAAPLT VPPVLRPASD
EVTRETEIAL RPTWVRLHPQ LPPTLMWGYD GQVPGPTIEV RRGQRVRIAW TNRIPKGSEY
PVTSVEVPLG PPGTPAPNTE PGRGGVEPNK DVAALPAWSV THLHGAQTGG GNDGWADNAV
GFGDAQLSEY PNDHQATQWW YHDHAMNITR WNVMAGLYGT YLVRDDEEDA LGLPSGDREI
PLLIADRNLD TDEDGRLNGR LLHKTVIVQQ SNPETGKPVS IPFFGPYTTV NGRIWPYADV
DDGWYRLRLV NASNARIYNL VLIDEDDRPV PGVVHQIGSD GGLLPRPVPV DFDDTLPVLS
AAPAERFDLL VDFRALGGRR LRLVDKGPGA PAGTPDPLGG VRYPEVMEFR VRETCEEDSF
ALPEVLSGSF RRMSHDIPHG HRLIVLTPPG TKGSGGHPEI WEMAEVEDPA DVQVPAEGVI
QVTGADGRTK TYRRTAATFN DGLGFTIGEG THEQWTFLNL SPILHPMHIH LADFQVLGRD
AYDASGFDLA LGGTRTPVRL DPDTPVPLAP NELGHKDVFQ VPGPQGLRVM GKFDGAYGRF
MYHCHLLEHE DMGMMRPFVV MPPEALKFDH GGAHGGHGEG HTG
