GCOB_AMYS7
ID GCOB_AMYS7 Reviewed; 334 AA.
AC P0DPQ8;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 10-OCT-2018, sequence version 1.
DT 03-AUG-2022, entry version 10.
DE RecName: Full=Aromatic O-demethylase, reductase subunit {ECO:0000305|PubMed:29950589};
DE EC=1.6.2.- {ECO:0000269|PubMed:29950589};
DE AltName: Full=NADH--hemoprotein reductase {ECO:0000305|PubMed:29950589};
GN Name=gcoB {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 (1.72 ANGSTROMS) IN COMPLEX WITH FAD AND 2FE-2S
RP CLUSTER, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBUNIT, AND DOMAIN.
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. GcoB transfers electrons from NADH to the cytochrome P450
CC subunit GcoA. Highly prefers NADH over NADPH as the electron donor.
CC {ECO:0000269|PubMed:29950589}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NADH + 2 oxidized [cytochrome P450] = H(+) + NAD(+) + 2
CC reduced [cytochrome P450]; Xref=Rhea:RHEA:57420, Rhea:RHEA-
CC COMP:14627, Rhea:RHEA-COMP:14628, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:55376, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945,
CC ChEBI:CHEBI:60344; Evidence={ECO:0000269|PubMed:29950589};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:29950589};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:29950589};
CC -!- COFACTOR:
CC Name=[2Fe-2S] cluster; Xref=ChEBI:CHEBI:190135;
CC Evidence={ECO:0000269|PubMed:29950589};
CC Note=Binds 1 [2Fe-2S] cluster per subunit.
CC {ECO:0000269|PubMed:29950589};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=16 uM for NADH {ECO:0000269|PubMed:29950589};
CC Note=kcat is 44 sec(-1) for NADH-mediated reduction of cytochrome c.
CC {ECO:0000269|PubMed:29950589};
CC -!- PATHWAY: Aromatic compound metabolism. {ECO:0000305|PubMed:29950589}.
CC -!- SUBUNIT: Monomer. Forms a heterodimer with GcoA.
CC {ECO:0000269|PubMed:29950589}.
CC -!- DOMAIN: GcoB has an unusual three-domain structure, with an N-terminal
CC 2Fe-2S domain and a C-terminal region that consists of an FAD-binding
CC domain followed by an NADH-binding domain.
CC {ECO:0000269|PubMed:29950589}.
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DR EMBL; AFWY03000000; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR RefSeq; WP_020419854.1; NZ_AFWY03000022.1.
DR PDB; 5OGX; X-ray; 1.72 A; A=1-334.
DR PDBsum; 5OGX; -.
DR AlphaFoldDB; P0DPQ8; -.
DR SMR; P0DPQ8; -.
DR GO; GO:0051537; F:2 iron, 2 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-KW.
DR CDD; cd00207; fer2; 1.
DR Gene3D; 3.10.20.30; -; 1.
DR Gene3D; 3.40.50.80; -; 1.
DR InterPro; IPR036010; 2Fe-2S_ferredoxin-like_sf.
DR InterPro; IPR001041; 2Fe-2S_ferredoxin-type.
DR InterPro; IPR006058; 2Fe2S_fd_BS.
DR InterPro; IPR012675; Beta-grasp_dom_sf.
DR InterPro; IPR008333; Cbr1-like_FAD-bd_dom.
DR InterPro; IPR017927; FAD-bd_FR_type.
DR InterPro; IPR001709; Flavoprot_Pyr_Nucl_cyt_Rdtase.
DR InterPro; IPR039261; FNR_nucleotide-bd.
DR InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR Pfam; PF00970; FAD_binding_6; 1.
DR Pfam; PF00111; Fer2; 1.
DR Pfam; PF00175; NAD_binding_1; 1.
DR PRINTS; PR00371; FPNCR.
DR SUPFAM; SSF52343; SSF52343; 1.
DR SUPFAM; SSF54292; SSF54292; 1.
DR SUPFAM; SSF63380; SSF63380; 1.
DR PROSITE; PS00197; 2FE2S_FER_1; 1.
DR PROSITE; PS51085; 2FE2S_FER_2; 1.
DR PROSITE; PS51384; FAD_FR; 1.
PE 1: Evidence at protein level;
KW 2Fe-2S; 3D-structure; Aromatic hydrocarbons catabolism; Electron transport;
KW FAD; Flavoprotein; Iron; Iron-sulfur; Metal-binding; NAD; Oxidoreductase;
KW Transport.
FT CHAIN 1..334
FT /note="Aromatic O-demethylase, reductase subunit"
FT /id="PRO_0000445480"
FT DOMAIN 1..91
FT /note="2Fe-2S ferredoxin-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT DOMAIN 98..198
FT /note="FAD-binding FR-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT BINDING 35
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 40
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 43
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 75
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 145..148
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 162..164
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 170..172
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 215
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 330
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29950589"
FT BINDING 334
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29950589"
SQ SEQUENCE 334 AA; 35566 MW; 176005B2CFFF7C7E CRC64;
MTFAVSVGGR RVDCEPGQTL LEAFLRGGVW MPNSCNQGTC GTCKLQVLSG EVDHGGAPED
TLSAEERASG LALACQARPL ADTEVRSTAD AGRVTHPLRD LTATVLEVAD IARDTRRVLL
GLAEPLAFEA GQYVELVVPG SGARRQYSLA NTADEDKVLE LHVRRVPGGV ATDGWLFDGL
AAGDRVEATG PLGDFHLPPP DEDDGGPMVL IGGGTGLAPL VGIARTALAR HPSREVLLYH
GVRGAADLYD LGRFAEIAEE HPGFRFVPVL SDEPDPAYRG GFPTDAFVED VPSGRGWSGW
LCGPPAMVEA GVKAFKRRRM SPRRIHREKF TPAS
