GAO_BARSP
ID GAO_BARSP Reviewed; 496 AA.
AC D5JBX1;
DT 21-SEP-2011, integrated into UniProtKB/Swiss-Prot.
DT 15-JUN-2010, sequence version 1.
DT 03-AUG-2022, entry version 42.
DE RecName: Full=Germacrene A hydroxylase {ECO:0000303|PubMed:20351109};
DE EC=1.14.14.95 {ECO:0000269|PubMed:20351109};
DE AltName: Full=Germacrene A oxidase {ECO:0000303|PubMed:20351109};
DE Short=BsGAO {ECO:0000303|PubMed:20351109};
GN Name=GAO {ECO:0000303|PubMed:20351109};
OS Barnadesia spinosa (Spiny barnadesia).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; campanulids; Asterales; Asteraceae; Barnadesioideae; Barnadesia.
OX NCBI_TaxID=171760;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=20351109; DOI=10.1074/jbc.m110.111757;
RA Nguyen D.T., Goepfert J.C., Ikezawa N., Macnevin G., Kathiresan M.,
RA Conrad J., Spring O., Ro D.-K.;
RT "Biochemical conservation and evolution of germacrene A oxidase in
RT asteraceae.";
RL J. Biol. Chem. 285:16588-16598(2010).
RN [2]
RP PATHWAY, AND REVIEW.
RX PubMed=30468448; DOI=10.1039/c8np00077h;
RA Liu Y., Jing S.-X., Luo S.-H., Li S.-H.;
RT "Non-volatile natural products in plant glandular trichomes: chemistry,
RT biological activities and biosynthesis.";
RL Nat. Prod. Rep. 36:626-665(2019).
CC -!- FUNCTION: Involved in the biosynthesis of germacrene-derived
CC sesquiterpene lactones (PubMed:20351109). Catalyzes three consecutive
CC oxidations of germacrene A to produce germacrene A acid
CC (PubMed:20351109). Could also catalyze the three-step oxidation of non-
CC natural substrate amorphadiene to artemisinic acid (PubMed:20351109).
CC {ECO:0000269|PubMed:20351109}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(+)-(R)-germacrene A + 3 O2 + 3 reduced [NADPH--hemoprotein
CC reductase] = germacra-1(10),4,11(13)-trien-12-oate + 4 H(+) + 4 H2O +
CC 3 oxidized [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:30303,
CC Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:41595,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:61301;
CC EC=1.14.14.95; Evidence={ECO:0000269|PubMed:20351109};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:30304;
CC Evidence={ECO:0000269|PubMed:20351109};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000303|PubMed:30468448}.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000250|UniProtKB:D5JBW8}; Single-pass type II membrane protein
CC {ECO:0000250|UniProtKB:D5JBW8}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; GU256647; ADF43083.1; -; mRNA.
DR AlphaFoldDB; D5JBX1; -.
DR SMR; D5JBX1; -.
DR BioCyc; MetaCyc:MON-15756; -.
DR BRENDA; 1.14.14.95; 12601.
DR UniPathway; UPA00213; -.
DR GO; GO:0005789; C:endoplasmic reticulum membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0106223; F:germacrene A hydroxylase activity; IDA:UniProtKB.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0051762; P:sesquiterpene biosynthetic process; IDA:UniProtKB.
DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Endoplasmic reticulum; Glycoprotein; Heme; Iron; Membrane; Metal-binding;
KW Monooxygenase; Oxidoreductase; Signal-anchor; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..496
FT /note="Germacrene A hydroxylase"
FT /id="PRO_0000412764"
FT TOPO_DOM 1..2
FT /note="Cytoplasmic"
FT /evidence="ECO:0000255"
FT TRANSMEM 3..23
FT /note="Helical; Signal-anchor for type II membrane protein"
FT /evidence="ECO:0000255"
FT TOPO_DOM 24..496
FT /note="Lumenal"
FT /evidence="ECO:0000255"
FT BINDING 432
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:0000250|UniProtKB:P04798"
FT CARBOHYD 493
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
SQ SEQUENCE 496 AA; 55699 MW; 028C2D4B45ED6909 CRC64;
MELTLTTSLG LAVFVFILFK LLTGSKSTKN SLPEAWRLPI IGHMHHLVGT LPHRGVTDMA
RKYGSLMHLQ LGEVSTIVVS SPRWAKEVLT TYDITFANRP ETLTGEIVAY HNTDIVLSPY
GEYWRQLRKL CTLELLSAKK VKSFQSLREE ECWNLVKEVR SSGSGSPVDL SESIFKLIAT
ILSRAAFGKG IKDQREFTEI VKEILRLTGG FDVADIFPSK KILHHLSGKR AKLTNIHNKL
DSLINNIVSE HPGSRTSSSQ ESLLDVLLRL KDSAELPLTS DNVKAVILDM FGAGTDTSSA
TIEWAISELI RCPRAMEKVQ TELRQALNGK ERIQEEDIQE LSYLKLVIKE TLRLHPPLPL
VMPRECREPC VLAGYEIPTK TKLIVNVFAI NRDPEYWKDA ETFMPERFEN SPINIMGSEY
EYLPFGAGRR MCPGAALGLA NVELPLAHIL YYFNWKLPNG ARLDELDMSE CFGATVQRKS
ELLLVPTAYK TANNSA
