C81E9_MEDTR
ID C81E9_MEDTR Reviewed; 501 AA.
AC Q6WNQ9;
DT 29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=Isoflavone 3'-hydroxylase {ECO:0000303|PubMed:14617078};
DE EC=1.14.14.88 {ECO:0000269|PubMed:14617078};
DE AltName: Full=Cytochrome P450 81E9 {ECO:0000303|PubMed:14617078};
DE Flags: Fragment;
GN Name=CYP81E9 {ECO:0000303|PubMed:14617078};
OS Medicago truncatula (Barrel medic) (Medicago tribuloides).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC NPAAA clade; Hologalegina; IRL clade; Trifolieae; Medicago.
OX NCBI_TaxID=3880 {ECO:0000312|EMBL:AAQ20041.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBCELLULAR LOCATION, TISSUE SPECIFICITY,
RP AND INDUCTION.
RX PubMed=14617078; DOI=10.1046/j.1365-313x.2003.01893.x;
RA Liu C.J., Huhman D., Sumner L.W., Dixon R.A.;
RT "Regiospecific hydroxylation of isoflavones by cytochrome P450 81E enzymes
RT from Medicago truncatula.";
RL Plant J. 36:471-484(2003).
RN [2]
RP IDENTIFICATION.
RX PubMed=24826896; DOI=10.1371/journal.pone.0097250;
RA Shearer A.G., Altman T., Rhee C.D.;
RT "Finding sequences for over 270 orphan enzymes.";
RL PLoS ONE 9:E97250-E97250(2014).
CC -!- FUNCTION: Involved in the biosynthesis of the pterocarpin phytoalexins.
CC Acts on isoflavones with a 4'-methoxy group on the B-ring, such as
CC biochanin A, formononetin and 2'-hydroxyformononetin. Has a low
CC activity with daidzein and pseudobaptigenin, and no activity with the
CC 7-O-methylated isoflavonoids isoformononetin and prunetin.
CC {ECO:0000269|PubMed:14617078}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=formononetin + O2 + reduced [NADPH--hemoprotein reductase] =
CC calycosin + H(+) + H2O + oxidized [NADPH--hemoprotein reductase];
CC Xref=Rhea:RHEA:22960, Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:77688,
CC ChEBI:CHEBI:77992; EC=1.14.14.88;
CC Evidence={ECO:0000269|PubMed:14617078};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:P04798};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=49.7 uM for formononetin {ECO:0000269|PubMed:14617078};
CC KM=112.6 uM for biochanin A {ECO:0000269|PubMed:14617078};
CC Note=kcat is 0.028 sec(-1) with formononetin as substrate. kcat is
CC 0.1 sec(-1) with biochanin A as substrate.
CC {ECO:0000269|PubMed:14617078};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:14617078};
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000269|PubMed:14617078}; Single-pass membrane protein
CC {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Expressed constitutively in leaves and stems, but
CC not in roots. {ECO:0000269|PubMed:14617078}.
CC -!- INDUCTION: Up-regulated by methyl jasmonate and herbivory, but only
CC weakly by elicitor. {ECO:0000269|PubMed:14617078}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family.
CC {ECO:0000255|RuleBase:RU000461}.
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DR EMBL; AY278228; AAQ20041.1; -; mRNA.
DR AlphaFoldDB; Q6WNQ9; -.
DR SMR; Q6WNQ9; -.
DR SABIO-RK; Q6WNQ9; -.
DR ExpressionAtlas; Q6WNQ9; differential.
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:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0048000; F:isoflavone 3'-hydroxylase activity; IEA:UniProtKB-EC.
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; Heme; Iron; Membrane; Metal-binding; Monooxygenase;
KW Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN <1..501
FT /note="Isoflavone 3'-hydroxylase"
FT /id="PRO_0000430744"
FT TRANSMEM 7..24
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 439
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 NON_TER 1
FT /evidence="ECO:0000312|EMBL:AAQ20041.1"
SQ SEQUENCE 501 AA; 57156 MW; B73CBBA506D54CCF CRC64;
ALFYYSLLSL SFIITIKILL KITSRRLKNL PPGPPTIPII GNLHHLKHPL HRTFTTLSQT
YGDIFSLWFG SRLVVVVSSP SLAHECFTKN DIILANRPRF LTGKYIFYNY TTLGSASYGD
HWRNLRRITT IDVLSNNRLN SFLGVRRDET NRLIQKLLKD VVSEGFGFTK VELRPRLTEM
TFNAMMRMIS GKRYYGDDGD VSDVEEAKQF REIISEMMSL LGANNKGDFL PLLRVVDLDN
LEKRCKRIAK RSNAFLEGLI EEHRRGNIHS DGGTMIDHLL KLSESQPEYY SDHLIKGLIQ
GMLLAGTDTS AVTIEWVMSE LLNHPEVLKK AKEELDTQIG KNKLVDEQDL SKLPYLQNII
SETLRLHPPA PLLLPHYSSE DCTIGEFNVP KDTIILTNVW GIHRDPKHWN DALSFKPERF
EKEEEVNKVM AFGLGRRACP GLSLAQRTVG FTVGLLIQCF EWERESEEKL DMMEGKGITM
PMKIPLRAMC KALPIANDVT K
