GCOA_AMYS7
ID GCOA_AMYS7 Reviewed; 407 AA.
AC P0DPQ7;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 10-OCT-2018, sequence version 1.
DT 03-AUG-2022, entry version 15.
DE RecName: Full=Aromatic O-demethylase, cytochrome P450 subunit {ECO:0000305|PubMed:29950589};
DE EC=1.14.14.- {ECO:0000269|PubMed:29950589};
GN Name=gcoA {ECO:0000303|PubMed:29950589};
OS Amycolatopsis sp. (strain ATCC 39116 / 75iv2).
OC Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae;
OC Amycolatopsis; unclassified Amycolatopsis.
OX NCBI_TaxID=385957;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 39116 / 75iv2;
RX PubMed=22493203; DOI=10.1128/jb.00186-12;
RA Davis J.R., Goodwin L.A., Woyke T., Teshima H., Bruce D., Detter C.,
RA Tapia R., Han S., Han J., Pitluck S., Nolan M., Mikhailova N., Land M.L.,
RA Sello J.K.;
RT "Genome sequence of Amycolatopsis sp. strain ATCC 39116, a plant biomass-
RT degrading actinomycete.";
RL J. Bacteriol. 194:2396-2397(2012).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.44 ANGSTROMS) IN COMPLEXES WITH HEME; GUAIACOL;
RP GUAETHOL; SYRINGOL AND VANILLIN, FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=ATCC 39116 / 75iv2;
RX PubMed=29950589; DOI=10.1038/s41467-018-04878-2;
RA Mallinson S.J.B., Machovina M.M., Silveira R.L., Garcia-Borras M.,
RA Gallup N., Johnson C.W., Allen M.D., Skaf M.S., Crowley M.F., Neidle E.L.,
RA Houk K.N., Beckham G.T., DuBois J.L., McGeehan J.E.;
RT "A promiscuous cytochrome P450 aromatic O-demethylase for lignin
RT bioconversion.";
RL Nat. Commun. 9:2487-2487(2018).
CC -!- FUNCTION: Part of a two-component P450 system that efficiently O-
CC demethylates diverse aromatic substrates such as guaiacol and a wide
CC variety of lignin-derived monomers. Is likely involved in lignin
CC degradation, allowing Amycolatopsis sp. ATCC 39116 to catabolize plant
CC biomass. GcoA binds and processes the substrate with electrons supplied
CC by the GcoB subunit. {ECO:0000269|PubMed:29950589}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=guaiacol + O2 + reduced [NADH--hemoprotein reductase] =
CC catechol + formaldehyde + H(+) + H2O + oxidized [NADH--hemoprotein
CC reductase]; Xref=Rhea:RHEA:57424, Rhea:RHEA-COMP:14893, Rhea:RHEA-
CC COMP:14894, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16842, ChEBI:CHEBI:18135, ChEBI:CHEBI:28591,
CC ChEBI:CHEBI:57692, ChEBI:CHEBI:58307;
CC Evidence={ECO:0000269|PubMed:29950589};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-methoxycatechol + O2 + reduced [NADH--hemoprotein reductase]
CC = 1,2,3-trihydroxybenzene + formaldehyde + H(+) + H2O + oxidized
CC [NADH--hemoprotein reductase]; Xref=Rhea:RHEA:57432, Rhea:RHEA-
CC COMP:14893, Rhea:RHEA-COMP:14894, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16164,
CC ChEBI:CHEBI:16842, ChEBI:CHEBI:57692, ChEBI:CHEBI:58307,
CC ChEBI:CHEBI:141700; Evidence={ECO:0000269|PubMed:29950589};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=anisole + O2 + reduced [NADH--hemoprotein reductase] =
CC formaldehyde + H(+) + H2O + oxidized [NADH--hemoprotein reductase] +
CC phenol; Xref=Rhea:RHEA:57428, Rhea:RHEA-COMP:14893, Rhea:RHEA-
CC COMP:14894, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:15882, ChEBI:CHEBI:16579, ChEBI:CHEBI:16842,
CC ChEBI:CHEBI:57692, ChEBI:CHEBI:58307;
CC Evidence={ECO:0000269|PubMed:29950589};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=guaethol + O2 + reduced [NADH--hemoprotein reductase] =
CC acetaldehyde + catechol + H(+) + H2O + oxidized [NADH--hemoprotein
CC reductase]; Xref=Rhea:RHEA:57436, Rhea:RHEA-COMP:14893, Rhea:RHEA-
CC COMP:14894, ChEBI:CHEBI:15343, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:18135, ChEBI:CHEBI:57692,
CC ChEBI:CHEBI:58307, ChEBI:CHEBI:141701;
CC Evidence={ECO:0000269|PubMed:29950589};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-methylanisole + O2 + reduced [NADH--hemoprotein reductase] =
CC 2-hydroxytoluene + formaldehyde + H(+) + H2O + oxidized [NADH--
CC hemoprotein reductase]; Xref=Rhea:RHEA:57440, Rhea:RHEA-COMP:14893,
CC Rhea:RHEA-COMP:14894, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16842, ChEBI:CHEBI:28054,
CC ChEBI:CHEBI:57692, ChEBI:CHEBI:58307, ChEBI:CHEBI:141702;
CC Evidence={ECO:0000269|PubMed:29950589};
CC -!- COFACTOR:
CC Name=heme b; Xref=ChEBI:CHEBI:60344;
CC Evidence={ECO:0000269|PubMed:29950589};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=60 uM for guaiacol {ECO:0000269|PubMed:29950589};
CC KM=30 uM for 3-methoxycatechol {ECO:0000269|PubMed:29950589};
CC KM=43 uM for anisole {ECO:0000269|PubMed:29950589};
CC KM=15 uM for guaethol {ECO:0000269|PubMed:29950589};
CC KM=27 uM for 2-methylanisole {ECO:0000269|PubMed:29950589};
CC Note=kcat is 6.8 sec(-1) with guaiacol as substrate. kcat is 2.1
CC sec(-1) with 3-methoxycatechol as substrate. kcat is 3.5 sec(-1) with
CC anisole as substrate. kcat is 1.4 sec(-1) with guaethol as substrate.
CC kcat is 4.6 sec(-1) with 2-methylanisole as substrate.
CC {ECO:0000269|PubMed:29950589};
CC -!- PATHWAY: Aromatic compound metabolism. {ECO:0000305|PubMed:29950589}.
CC -!- SUBUNIT: Monomer. Forms a heterodimer with GcoB.
CC {ECO:0000269|PubMed:29950589}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; AFWY03000000; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR RefSeq; WP_020419855.1; NZ_AFWY03000022.1.
DR PDB; 5NCB; X-ray; 1.44 A; A=1-407.
DR PDB; 5OMR; X-ray; 1.68 A; A=1-407.
DR PDB; 5OMS; X-ray; 1.95 A; A=1-407.
DR PDB; 5OMU; X-ray; 1.95 A; A=1-407.
DR PDB; 6HQK; X-ray; 1.57 A; A=1-407.
DR PDB; 6HQL; X-ray; 1.49 A; A=1-407.
DR PDB; 6HQM; X-ray; 1.85 A; A=1-407.
DR PDB; 6HQN; X-ray; 1.87 A; A=1-407.
DR PDB; 6HQO; X-ray; 1.70 A; A=1-407.
DR PDB; 6HQP; X-ray; 1.62 A; A=1-407.
DR PDB; 6HQQ; X-ray; 1.66 A; A=1-407.
DR PDB; 6HQR; X-ray; 1.79 A; A=1-407.
DR PDB; 6HQS; X-ray; 2.17 A; A=1-407.
DR PDB; 6HQT; X-ray; 1.85 A; A=1-407.
DR PDB; 6YCH; X-ray; 1.88 A; A=1-407.
DR PDB; 6YCI; X-ray; 1.80 A; A=1-407.
DR PDB; 6YCJ; X-ray; 1.64 A; A=1-407.
DR PDB; 6YCK; X-ray; 1.80 A; A=1-407.
DR PDB; 6YCL; X-ray; 1.70 A; A=1-407.
DR PDB; 6YCM; X-ray; 1.60 A; A=1-407.
DR PDB; 6YCN; X-ray; 1.83 A; A=1-407.
DR PDB; 6YCO; X-ray; 1.80 A; A=1-407.
DR PDB; 6YCP; X-ray; 1.80 A; A=1-407.
DR PDB; 6YCT; X-ray; 2.39 A; A=1-407.
DR PDBsum; 5NCB; -.
DR PDBsum; 5OMR; -.
DR PDBsum; 5OMS; -.
DR PDBsum; 5OMU; -.
DR PDBsum; 6HQK; -.
DR PDBsum; 6HQL; -.
DR PDBsum; 6HQM; -.
DR PDBsum; 6HQN; -.
DR PDBsum; 6HQO; -.
DR PDBsum; 6HQP; -.
DR PDBsum; 6HQQ; -.
DR PDBsum; 6HQR; -.
DR PDBsum; 6HQS; -.
DR PDBsum; 6HQT; -.
DR PDBsum; 6YCH; -.
DR PDBsum; 6YCI; -.
DR PDBsum; 6YCJ; -.
DR PDBsum; 6YCK; -.
DR PDBsum; 6YCL; -.
DR PDBsum; 6YCM; -.
DR PDBsum; 6YCN; -.
DR PDBsum; 6YCO; -.
DR PDBsum; 6YCP; -.
DR PDBsum; 6YCT; -.
DR AlphaFoldDB; P0DPQ7; -.
DR SMR; P0DPQ7; -.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro.
DR GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR002397; Cyt_P450_B.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00359; BP450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aromatic hydrocarbons catabolism; Heme; Iron; Metal-binding;
KW Monooxygenase; Oxidoreductase.
FT CHAIN 1..407
FT /note="Aromatic O-demethylase, cytochrome P450 subunit"
FT /id="PRO_0000445479"
FT BINDING 88
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 92
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 241
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 245
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 298
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 321
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 351
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 354
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 356
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000269|PubMed:29950589"
FT HELIX 7..14
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 17..22
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 25..34
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 36..40
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 41..43
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 45..48
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 51..59
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 63..66
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 69..75
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 80..82
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 85..99
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 101..121
FT /evidence="ECO:0007829|PDB:6HQL"
FT TURN 122..125
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 126..130
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 131..134
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 136..148
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 155..169
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 178..181
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 183..186
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 187..209
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 215..221
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 232..263
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 265..273
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 275..277
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 278..288
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 291..294
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 296..302
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 304..306
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 309..311
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 316..319
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 321..324
FT /evidence="ECO:0007829|PDB:6HQL"
FT TURN 328..330
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 331..333
FT /evidence="ECO:0007829|PDB:6YCH"
FT HELIX 352..354
FT /evidence="ECO:0007829|PDB:6HQL"
FT HELIX 359..376
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 384..386
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 393..395
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 397..399
FT /evidence="ECO:0007829|PDB:6HQL"
FT STRAND 402..404
FT /evidence="ECO:0007829|PDB:6HQL"
SQ SEQUENCE 407 AA; 45253 MW; 41EC44A2FA3DAFFF CRC64;
MTTTERPDLA WLDEVTMTQL ERNPYEVYER LRAEAPLAFV PVLGSYVAST AEVCREVATS
PDFEAVITPA GGRTFGHPAI IGVNGDIHAD LRSMVEPALQ PAEVDRWIDD LVRPIARRYL
ERFENDGHAE LVAQYCEPVS VRSLGDLLGL QEVDSDKLRE WFAKLNRSFT NAAVDENGEF
ANPEGFAEGD QAKAEIRAVV DPLIDKWIEH PDDSAISHWL HDGMPPGQTR DREYIYPTIY
VYLLGAMQEP GHGMASTLVG LFSRPEQLEE VVDDPTLIPR AIAEGLRWTS PIWSATARIS
TKPVTIAGVD LPAGTPVMLS YGSANHDTGK YEAPSQYDLH RPPLPHLAFG AGNHACAGIY
FANHVMRIAL EELFEAIPNL ERDTREGVEF WGWGFRGPTS LHVTWEV
