C71Z4_PASSA
ID C71Z4_PASSA Reviewed; 504 AA.
AC A0A2Z5D854;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 10-OCT-2018, sequence version 1.
DT 03-AUG-2022, entry version 10.
DE RecName: Full=Xanthotoxol synthase {ECO:0000303|PubMed:29971079};
DE EC=1.14.14.- {ECO:0000269|PubMed:29971079};
DE AltName: Full=Cytochrome P450 CYP71AZ4 {ECO:0000303|PubMed:29971079};
DE AltName: Full=Fraxetin synthase {ECO:0000303|PubMed:29971079};
DE EC=1.14.14.- {ECO:0000269|PubMed:29971079};
GN Name=CYP71AZ4 {ECO:0000303|PubMed:29971079};
OS Pastinaca sativa (Wild parsnip) (Anethum pastinaca).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; campanulids; Apiales; Apiaceae; Apioideae; apioid superclade;
OC Tordylieae; Tordyliinae; Pastinaca.
OX NCBI_TaxID=4041;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, MUTAGENESIS OF 108-GLY--VAL-129;
RP 280-LYS--PHE-310 AND 364-GLY--ALA-372, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, REVIEW, PATHWAY, AND INDUCTION BY WOUNDING.
RC STRAIN=cv. Demi-long de Guernesey;
RX PubMed=29971079; DOI=10.3389/fpls.2018.00820;
RA Krieger C., Roselli S., Kellner-Thielmann S., Galati G., Schneider B.,
RA Grosjean J., Olry A., Ritchie D., Matern U., Bourgaud F., Hehn A.;
RT "The CYP71AZ P450 subfamily: A driving factor for the diversification of
RT coumarin biosynthesis in apiaceous plants.";
RL Front. Plant Sci. 9:820-820(2018).
CC -!- FUNCTION: Involved in the biosynthesis of coumarins and furanocoumarins
CC (FCs), natural products required for defense responses against attacks
CC by predators with potential medical and agroindustrial usages such as
CC anticoagulant, rodenticide and artificial vanilla substitutes
CC (PubMed:29971079). Catalyzes the conversion of psoralen into
CC xanthotoxol and of 6-methoxycoumarin into scopoletin (PubMed:29971079).
CC Can also convert with a lower efficiency scopoletin into fraxetin and
CC 7-methoxycoumarin into daphnetin-7-methylether, and use 7-methoxy-3-
CC methylcoumarin as substrate (PubMed:29971079).
CC {ECO:0000269|PubMed:29971079, ECO:0000303|PubMed:29971079}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=O2 + psoralen + reduced [NADPH--hemoprotein reductase] = H(+)
CC + H2O + oxidized [NADPH--hemoprotein reductase] + xanthotoxol;
CC Xref=Rhea:RHEA:68548, Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:15709, ChEBI:CHEBI:27616, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210; Evidence={ECO:0000269|PubMed:29971079};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68549;
CC Evidence={ECO:0000269|PubMed:29971079};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=6-methoxycoumarin + O2 + reduced [NADPH--hemoprotein
CC reductase] = H(+) + H2O + oxidized [NADPH--hemoprotein reductase] +
CC scopoletin; Xref=Rhea:RHEA:68564, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17488, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC ChEBI:CHEBI:178005; Evidence={ECO:0000269|PubMed:29971079};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68565;
CC Evidence={ECO:0000269|PubMed:29971079};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q94IP1};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=11.3 uM for psoralen (at pH 7.4 and 28 degrees Celsius)
CC {ECO:0000269|PubMed:29971079};
CC KM=9.5 uM for 6-methoxycoumarin (at pH 7.4 and 28 degrees Celsius)
CC {ECO:0000269|PubMed:29971079};
CC KM=762.2 uM for scopoletin (at pH 7.4 and 28 degrees Celsius)
CC {ECO:0000269|PubMed:29971079};
CC KM=159.9 uM for 7-methoxy-3-methylcoumarin (at pH 7.4 and 28 degrees
CC Celsius) {ECO:0000269|PubMed:29971079};
CC KM=72.5 uM for 7-methoxycoumarin (at pH 7.4 and 28 degrees Celsius)
CC {ECO:0000269|PubMed:29971079};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:29971079}.
CC -!- SUBCELLULAR LOCATION: Microsome membrane
CC {ECO:0000250|UniProtKB:Q6QNI4}; Single-pass membrane protein
CC {ECO:0000255}.
CC -!- INDUCTION: Accumulates in roots after wounding.
CC {ECO:0000269|PubMed:29971079}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; MH000219; AXB38861.1; -; mRNA.
DR KEGG; ag:AXB38861; -.
DR GO; GO:0005783; C:endoplasmic reticulum; IEA:UniProtKB-KW.
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: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:0009805; P:coumarin biosynthetic process; IDA:UniProtKB.
DR GO; GO:0009611; P:response to wounding; IEP:UniProtKB.
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; Microsome;
KW Monooxygenase; Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..504
FT /note="Xanthotoxol synthase"
FT /id="PRO_0000454523"
FT TRANSMEM 3..23
FT /note="Helical"
FT /evidence="ECO:0000255"
FT REGION 363..368
FT /note="Substrate specificity"
FT /evidence="ECO:0000269|PubMed:29971079"
FT BINDING 444
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:Q94IP1"
FT SITE 118
FT /note="Substrate specificity"
FT /evidence="ECO:0000269|PubMed:29971079"
FT SITE 299
FT /note="Substrate specificity"
FT /evidence="ECO:0000269|PubMed:29971079"
FT SITE 303
FT /note="Substrate specificity"
FT /evidence="ECO:0000269|PubMed:29971079"
FT MUTAGEN 108..129
FT /note="GKLSYNGLEMAFAPYGEHWRNV->QKLSYNGLDVAFSPYTDQWRHT: In
FT Mut1; lost activity on fraxetin, scopoletin, 6-
FT methoxycoumarin, 7-methoxycoumarin and 7-methoxy-3-
FT methylcoumarin, but acquired activity on esculetin. In
FT Mut4; restricted substrate specificity to esculetin, thus
FT mimicking CYP71AZ3 activity; when associated with 280-E--W-
FT 312 and 364-A--T-372."
FT /evidence="ECO:0000269|PubMed:29971079"
FT MUTAGEN 280..310
FT /note="KGSFLTMDSIKAILLNVFSGGIGTTGSALVF->EGSSLTKDSMKAILLNVFN
FT GGTGTTATVLAW: In Mut2; lost activity on psoralen but
FT acquired activity on esculetin. In Mut4; restricted
FT substrate specificity to esculetin, thus mimicking CYP71AZ3
FT activity; when associated with 108-Q--T-129 and 364-A--T-
FT 372."
FT /evidence="ECO:0000269|PubMed:29971079"
FT MUTAGEN 364..372
FT /note="GPLLIPRVA->APVLVPRET: In Mut3; reduced activity on
FT 6-methoxycoumarin and 7-methoxy-3-methylcoumarin, but
FT acquired activity on esculetin. In Mut4; restricted
FT substrate specificity to esculetin, thus mimicking CYP71AZ3
FT activity; when associated with 108-Q--T-129 and 280-E--W-
FT 312."
FT /evidence="ECO:0000269|PubMed:29971079"
SQ SEQUENCE 504 AA; 57254 MW; F42BBE237ECB77BE CRC64;
MDPAAIFLIL AIPIASVYLL FYHKKRVNGL SSPPGPRGLP FIGHFYQIYK SECAHEYISN
LSKQYGSLMT LHLGSVPALV VSSPKMAQEV LKTQDLVFCS RAQMTGSGKL SYNGLEMAFA
PYGEHWRNVR KMCTLELFTQ KRAQFNFRPV REDEVSRMVG RLSEAAAASE DVNAYECFTN
FATSIISRVA FGKRYDEDNL GKEKFQRMVA DIEAMFAAFF VSDFFPMFGW IDRLSGVKAV
LDRNFNEMDT FYQELIDEHL KPDRPESLNG DLIDVMLKNK GSFLTMDSIK AILLNVFSGG
IGTTGSALVF AMTALLRNQR VMKKAQEEVR SVIGKKEIVD EDDIQKLPYL RAVVKETLRL
YPPGPLLIPR VAMESCVLGE DEDHMYMIKP NTIVYVNTWG IGRDPKYWKN PLEFMPERFF
ERPDLNYTGQ QFEYLPFGSG RRICAGIIIG QNNVEVGLAN LLYSFDWEPP TGKTFEDIDD
QPCNGLTLAK KNPLYIRPKI YVHP
