GRIF_STRGG
ID GRIF_STRGG Reviewed; 306 AA.
AC B1VTI5; Q4W5X1;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 20-MAY-2008, sequence version 1.
DT 03-AUG-2022, entry version 63.
DE RecName: Full=Grixazone synthase {ECO:0000303|PubMed:16282322};
DE EC=1.10.3.15 {ECO:0000269|PubMed:16282322};
DE AltName: Full=O-aminophenol oxidase {ECO:0000303|PubMed:16282322};
DE EC=1.10.3.4 {ECO:0000269|PubMed:16282322};
DE AltName: Full=Phenoxazinone synthase {ECO:0000303|PubMed:16282322};
DE Short=PHS {ECO:0000303|PubMed:16282322};
GN Name=griF {ECO:0000303|PubMed:16282322}; OrderedLocusNames=SGR_4246;
OS Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350).
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=455632;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP BIOPHYSICOCHEMICAL PROPERTIES, DISRUPTION PHENOTYPE, ACTIVITY REGULATION,
RP AND SUBSTRATE SPECIFICITY.
RC STRAIN=JCM 4626 / NBRC 13350;
RX PubMed=16282322; DOI=10.1074/jbc.m505806200;
RA Suzuki H., Furusho Y., Higashi T., Ohnishi Y., Horinouchi S.;
RT "A novel o-aminophenol oxidase responsible for formation of the
RT phenoxazinone chromophore of grixazone.";
RL J. Biol. Chem. 281:824-833(2006).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=JCM 4626 / NBRC 13350;
RX PubMed=18375553; DOI=10.1128/jb.00204-08;
RA Ohnishi Y., Ishikawa J., Hara H., Suzuki H., Ikenoya M., Ikeda H.,
RA Yamashita A., Hattori M., Horinouchi S.;
RT "Genome sequence of the streptomycin-producing microorganism Streptomyces
RT griseus IFO 13350.";
RL J. Bacteriol. 190:4050-4060(2008).
RN [3]
RP REVIEW, AND FUNCTION.
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).
CC -!- FUNCTION: Involved in the biosynthesis of the parasiticide antibiotic
CC grixazone. Catalyzes the oxidation of 3,4-aminohydroxybenzoate (3,4-
CC AHBOA) to yield the corresponding quinone imine which is then non-
CC enzymatically conjugated with the thiol group of N-acetylcysteine. The
CC resultant compound is oxidized to its quinone imine enzymatically and
CC is then dimerized non-enzymatically with another quinone imine oxidized
CC by GriF to yield grixazone B. 3,4-aminohydroxybenzaldehyde (3,4-AHBAL)
CC can also be used as substrate to yield grixazone A. In the grixazone
CC biosynthetic pathway, it can also function as an o-aminophenol oxidase
CC that catalyzes the formation of the phenoxazinone chromophore from
CC alpha-aminophenol. It can also use 3,4-dihydroxybenzaldehyde, 2-amino-
CC 4-methylphenol and 3,4-dihydroxy-L-phenylalanine (L-DOPA) as
CC substrates. In contrast to tyrosinases, it does not display
CC monophenolase activity. {ECO:0000269|PubMed:16282322,
CC ECO:0000303|PubMed:19268377}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 3-amino-4-hydroxybenzoate + H(+) + N-acetyl-L-cysteine + 2
CC O2 = CO2 + grixazone B + 4 H2O; Xref=Rhea:RHEA:41420,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:60005, ChEBI:CHEBI:73483,
CC ChEBI:CHEBI:78236; EC=1.10.3.15;
CC Evidence={ECO:0000269|PubMed:16282322};
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:16282322};
CC -!- COFACTOR:
CC Name=Cu(2+); Xref=ChEBI:CHEBI:29036;
CC Evidence={ECO:0000269|PubMed:16282322};
CC Note=Binds 2 copper ions per subunit. {ECO:0000269|PubMed:16282322};
CC -!- ACTIVITY REGULATION: Inhibited by 3-amino-4-hydroxybenzensulfonic acid,
CC 4-hydroxy-3-nitrobenzaldehyde, L-tyrosine, P-hydroxybenzaldehyde.
CC Activated by the copper chaperone GriE. {ECO:0000269|PubMed:16282322}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.5 mM for alpha-aminophenol {ECO:0000269|PubMed:16282322};
CC KM=0.58 mM for 3,4-AHBAL {ECO:0000269|PubMed:16282322};
CC KM=0.75 mM for 2-amino-4-methylphenol {ECO:0000269|PubMed:16282322};
CC KM=0.41 mM for 3,4-dihydroxybenzaldehyde
CC {ECO:0000269|PubMed:16282322};
CC KM=19 mM for catechol {ECO:0000269|PubMed:16282322};
CC KM=5.5 mM for L-DOPA {ECO:0000269|PubMed:16282322};
CC Note=kcat is 20 sec(-1) with alpha-aminophenol as substrate. kcat is
CC 14 sec(-1) with 3,4-AHBAL as substrate. kcat is 18 sec(-1) with 2-
CC amino-4-methylphenol as substrate. kcat is 0.8 sec(-1) with 3,4-
CC dihydroxybenzaldehyde as substrate. kcat is 12 sec(-1) with catechol
CC as substrate. kcat is 0.066 sec(-1) with L-DOPA as substrate.
CC {ECO:0000269|PubMed:16282322};
CC pH dependence:
CC Optimum pH is between 8.5 and 10.5. {ECO:0000269|PubMed:16282322};
CC Temperature dependence:
CC Optimum temperature is 55 degrees Celsius.
CC {ECO:0000269|PubMed:16282322};
CC -!- DISRUPTION PHENOTYPE: Cells lacking griE and griF show accumulation of
CC the 3-amino-4-hydroxybenzaldehyde (3,4-AHBAL) intermediate.
CC {ECO:0000269|PubMed:16282322}.
CC -!- SIMILARITY: Belongs to the tyrosinase family. {ECO:0000305}.
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DR EMBL; AB214954; BAD99129.1; -; Genomic_DNA.
DR EMBL; AB259663; BAF36648.1; -; Genomic_DNA.
DR EMBL; AP009493; BAG21075.1; -; Genomic_DNA.
DR RefSeq; WP_012380459.1; NC_010572.1.
DR AlphaFoldDB; B1VTI5; -.
DR SMR; B1VTI5; -.
DR STRING; 455632.SGR_4246; -.
DR EnsemblBacteria; BAG21075; BAG21075; SGR_4246.
DR GeneID; 6212571; -.
DR KEGG; sgr:SGR_4246; -.
DR PATRIC; fig|455632.4.peg.4326; -.
DR eggNOG; COG2304; Bacteria.
DR HOGENOM; CLU_035914_3_0_11; -.
DR OMA; WHRYFTK; -.
DR OrthoDB; 446922at2; -.
DR BioCyc; MetaCyc:MON-12101; -.
DR Proteomes; UP000001685; Chromosome.
DR GO; GO:0005507; F:copper ion binding; IDA:UniProtKB.
DR GO; GO:0050149; F:o-aminophenol oxidase activity; IDA:UniProtKB.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 1.10.1280.10; -; 1.
DR InterPro; IPR008922; Di-copper_centre_dom_sf.
DR InterPro; IPR002227; Tyrosinase_Cu-bd.
DR Pfam; PF00264; Tyrosinase; 1.
DR PRINTS; PR00092; TYROSINASE.
DR SUPFAM; SSF48056; SSF48056; 1.
DR PROSITE; PS00497; TYROSINASE_1; 1.
DR PROSITE; PS00498; TYROSINASE_2; 1.
PE 1: Evidence at protein level;
KW Antibiotic biosynthesis; Copper; Metal-binding; Oxidoreductase.
FT CHAIN 1..306
FT /note="Grixazone synthase"
FT /id="PRO_0000418500"
FT BINDING 39
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="A"
FT /evidence="ECO:0000305|PubMed:16282322,
FT ECO:0000305|PubMed:19268377"
FT BINDING 58
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="A"
FT /evidence="ECO:0000305|PubMed:16282322,
FT ECO:0000305|PubMed:19268377"
FT BINDING 67
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="A"
FT /evidence="ECO:0000305|PubMed:16282322,
FT ECO:0000305|PubMed:19268377"
FT BINDING 222
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="B"
FT /evidence="ECO:0000305|PubMed:16282322,
FT ECO:0000305|PubMed:19268377"
FT BINDING 226
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="B"
FT /evidence="ECO:0000305|PubMed:16282322,
FT ECO:0000305|PubMed:19268377"
FT BINDING 248
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="B"
FT /evidence="ECO:0000305|PubMed:16282322,
FT ECO:0000305|PubMed:19268377"
SQ SEQUENCE 306 AA; 35583 MW; 7AA0E45A33795EE5 CRC64;
MVHVRKNHLT MTAEEKRRFV HAVLEIKRRG IYDRFVKLHI QINSTDYLDK ETGKRLGHVN
PGFLPWHRQY LLKFEQALQK VDPRVTLPYW DWTTDHGENS PLWSDTFMGG NGRPGDRRVM
TGPFARRNGW KLNISVIPEG PEDPALNGNY THDDRDYLVR DFGTLTPDLP TPQELEQTLD
LTVYDCPPWN HTSGGTPPYE SFRNHLEGYT KFAWEPRLGK LHGAAHVWTG GHMMYIGSPN
DPVFFLNHCM IDRCWALWQA RHPDVPHYLP TVPTQDVPDL NTPLGPWHTK TPADLLDHTR
FYTYDQ
