GAO_CICIN
ID GAO_CICIN Reviewed; 488 AA.
AC D5JBW8;
DT 21-SEP-2011, integrated into UniProtKB/Swiss-Prot.
DT 15-JUN-2010, sequence version 1.
DT 03-AUG-2022, entry version 44.
DE RecName: Full=Germacrene A hydroxylase;
DE EC=1.14.14.95 {ECO:0000269|PubMed:11299372, ECO:0000269|PubMed:20351109};
DE AltName: Full=Germacrene A oxidase;
DE Short=CiGAO;
OS Cichorium intybus (Chicory).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; campanulids; Asterales; Asteraceae; Cichorioideae; Cichorieae;
OC Cichoriinae; Cichorium.
OX NCBI_TaxID=13427;
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 FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY
RP REGULATION.
RX PubMed=11299372; DOI=10.1104/pp.125.4.1930;
RA de Kraker J.W., Franssen M.C., Dalm M.C., de Groot A., Bouwmeester H.J.;
RT "Biosynthesis of germacrene A carboxylic acid in chicory roots.
RT Demonstration of a cytochrome P450 (+)-germacrene a hydroxylase and NADP+-
RT dependent sesquiterpenoid dehydrogenase(s) involved in sesquiterpene
RT lactone biosynthesis.";
RL Plant Physiol. 125:1930-1940(2001).
RN [3]
RP FUNCTION, AND SUBCELLULAR LOCATION.
RA de Kraker J.W., Schurink M., Franssen M.C., Koenig W.A., de Groot A.,
RA Bouwmeester H.J.;
RT "Hydroxylation of sesquiterpenes by enzymes from chicory (Cichorium intybus
RT L.) roots.";
RL Tetrahedron 59:409-418(2003).
RN [4]
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. Catalyzes three consecutive oxidations of
CC germacrene A to produce germacrene A acid. Could also catalyze the
CC three-step oxidation of non-natural substrate amorphadiene to
CC artemisinic acid. Can use beta-elemene as substrate.
CC {ECO:0000269|PubMed:11299372, ECO:0000269|PubMed:20351109,
CC ECO:0000269|Ref.3}.
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:11299372,
CC ECO:0000269|PubMed:20351109};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:30304;
CC Evidence={ECO:0000269|PubMed:11299372, ECO:0000269|PubMed:20351109};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC -!- ACTIVITY REGULATION: Inhibited by cytochrome C, miconazole,
CC aminobenzotriazole, metyrapone and clotrimazole.
CC {ECO:0000269|PubMed:11299372}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:11299372};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000303|PubMed:30468448}.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000269|Ref.3}; Single-pass type II membrane protein
CC {ECO:0000269|Ref.3}. Microsome membrane {ECO:0000269|Ref.3}; Single-
CC pass type II membrane protein {ECO:0000269|Ref.3}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; GU256644; ADF43080.1; -; mRNA.
DR AlphaFoldDB; D5JBW8; -.
DR SMR; D5JBW8; -.
DR PRIDE; D5JBW8; -.
DR KEGG; ag:ADF43080; -.
DR BioCyc; MetaCyc:MON-15752; -.
DR BRENDA; 1.14.14.95; 1385.
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 Microsome; Monooxygenase; Oxidoreductase; Signal-anchor; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..488
FT /note="Germacrene A hydroxylase"
FT /id="PRO_0000412762"
FT TOPO_DOM 1..6
FT /note="Cytoplasmic"
FT /evidence="ECO:0000255"
FT TRANSMEM 7..23
FT /note="Helical; Signal-anchor for type II membrane protein"
FT /evidence="ECO:0000255"
FT TOPO_DOM 24..488
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 260
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 379
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
SQ SEQUENCE 488 AA; 54972 MW; C80A0EA15D63D8BE CRC64;
MELSLTTSIA LATIVLILYK LATRPKSNKK RLPEASRLPI IGHMHHLIGT MPHRGVMELA
RKHGSLMHLQ LGEVSTIVVS SPKWAKEILT TYDITFANRP ETLTGEIIAY HNTDIVLAPY
GEYWRQLRKL CTLELLSVKK VKSFQSIREE ECWNLVKEVK ESGSGKPISL SESIFKMIAT
ILSRAAFGKG IKDQREFTEI VKEILRQTGG FDVADIFPSK KFLHHLSGKR ARLTSIHKKL
DTLINNIVAE HHVSTSSKAN ETLLDVLLRL KDSAEFPLTA DNVKAIILDM FGAGTDTSSA
TVEWAISELI RCPRAMEKVQ AELRQALNGK EQIHEEDIQD LPYLNLVIRE TLRLHPPLPL
VMPRECREPV NLAGYEIANK TKLIVNVFAI NRDPEYWKDA EAFIPERFEN NPNNIMGADY
EYLPFGAGRR MCPGAALGLA NVQLPLANIL YHFNWKLPNG ASHDQLDMTE SFGATVQRKT
ELILVPSF
