T3O_CATRO
ID T3O_CATRO Reviewed; 506 AA.
AC I1TEM1;
DT 10-MAY-2017, integrated into UniProtKB/Swiss-Prot.
DT 11-JUL-2012, sequence version 1.
DT 03-AUG-2022, entry version 38.
DE RecName: Full=Tabersonine 3-oxygenase {ECO:0000303|PubMed:25918424};
DE Short=T3O {ECO:0000303|PubMed:25918424};
DE EC=1.14.14.50 {ECO:0000269|PubMed:25918424};
DE AltName: Full=16-methoxytabersonine 3-oxygenase {ECO:0000303|PubMed:25850027};
DE AltName: Full=Cytochrome P450 71D1 {ECO:0000303|PubMed:25850027};
DE AltName: Full=Cytochrome P450 71D1V2 {ECO:0000303|PubMed:22837051};
GN Name=CYP71D1V2 {ECO:0000303|PubMed:22837051};
GN Synonyms=16T3O {ECO:0000303|PubMed:25850027},
GN CYP71D1 {ECO:0000303|PubMed:25850027}, T3O {ECO:0000303|PubMed:25918424};
OS Catharanthus roseus (Madagascar periwinkle) (Vinca rosea).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Gentianales; Apocynaceae; Rauvolfioideae; Vinceae;
OC Catharanthinae; Catharanthus.
OX NCBI_TaxID=4058 {ECO:0000312|EMBL:AEX07771.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], AND SUBSTRATE SPECIFICITY.
RX PubMed=22837051; DOI=10.1007/s00425-012-1712-0;
RA Huang L., Li J., Ye H., Li C., Wang H., Liu B., Zhang Y.;
RT "Molecular characterization of the pentacyclic triterpenoid biosynthetic
RT pathway in Catharanthus roseus.";
RL Planta 236:1571-1581(2012).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, TISSUE
RP SPECIFICITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=25918424; DOI=10.1073/pnas.1501821112;
RA Qu Y., Easson M.L., Froese J., Simionescu R., Hudlicky T., De Luca V.;
RT "Completion of the seven-step pathway from tabersonine to the anticancer
RT drug precursor vindoline and its assembly in yeast.";
RL Proc. Natl. Acad. Sci. U.S.A. 112:6224-6229(2015).
RN [3]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, AND SUBCELLULAR LOCATION.
RC STRAIN=cv. Little Bright Eyes;
RX PubMed=25850027; DOI=10.1039/c5cc01309g;
RA Kellner F., Geu-Flores F., Sherden N.H., Brown S., Foureau E.,
RA Courdavault V., O'Connor S.E.;
RT "Discovery of a P450-catalyzed step in vindoline biosynthesis: a link
RT between the aspidosperma and eburnamine alkaloids.";
RL Chem. Commun. (Camb.) 51:7626-7628(2015).
RN [4]
RP INDUCTION BY HERBIVORY.
RX PubMed=28094274; DOI=10.1038/srep40453;
RA Duge de Bernonville T., Carqueijeiro I., Lanoue A., Lafontaine F.,
RA Sanchez Bel P., Liesecke F., Musset K., Oudin A., Glevarec G., Pichon O.,
RA Besseau S., Clastre M., St-Pierre B., Flors V., Maury S., Huguet E.,
RA O'Connor S.E., Courdavault V.;
RT "Folivory elicits a strong defense reaction in Catharanthus roseus:
RT metabolomic and transcriptomic analyses reveal distinct local and systemic
RT responses.";
RL Sci. Rep. 7:40453-40453(2017).
CC -!- FUNCTION: Cytochrome P450 catalyzing the monooxygenation of 16-
CC methoxytabersonine, 16-hydroxytabersonine and tabersonine, but not of
CC 2,3-dihydrotabersonine (PubMed:25918424). Converts the C2,C3 alkene of
CC tabersonine and 16-methoxytabersonine to the epoxides, which then
CC spontaneously open to form the corresponding imine alcohols
CC (PubMed:25850027). Inactive in converting amyrin to ursolic acid
CC (PubMed:22837051). {ECO:0000269|PubMed:22837051,
CC ECO:0000269|PubMed:25850027, ECO:0000269|PubMed:25918424}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=16-methoxytabersonine + O2 + reduced [NADPH--hemoprotein
CC reductase] = (3R)-1,2-didehydro-3-hydroxy-16-methoxy-2,3-
CC dihydrotabersonine + H(+) + H2O + oxidized [NADPH--hemoprotein
CC reductase]; Xref=Rhea:RHEA:52480, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:58930,
CC ChEBI:CHEBI:136640; EC=1.14.14.50;
CC Evidence={ECO:0000269|PubMed:25918424};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(-)-tabersonine + O2 + reduced [NADPH--hemoprotein reductase]
CC = (3R)-1,2-didehydro-3-hydroxy-2,3-dihydrotabersonine + H(+) + H2O +
CC oxidized [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:55024,
CC Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:57893, ChEBI:CHEBI:58210, ChEBI:CHEBI:138462;
CC EC=1.14.14.50; Evidence={ECO:0000269|PubMed:25918424};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q96242};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.1 uM for tabersonine {ECO:0000269|PubMed:25918424};
CC Note=The KM is mesured in the presence of a saturating amount of
CC tabersonine 3-reductase, the enzyme converting the unstable
CC intermediate produced by the reaction. {ECO:0000269|PubMed:25918424};
CC -!- PATHWAY: Alkaloid biosynthesis; vindoline biosynthesis.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000269|PubMed:25850027}; Single-pass membrane protein.
CC -!- TISSUE SPECIFICITY: Expressed in leaf epidermis.
CC {ECO:0000269|PubMed:25918424}.
CC -!- INDUCTION: Up-regulated by herbivory. {ECO:0000269|PubMed:28094274}.
CC -!- MISCELLANEOUS: The CYP71D1V2 product is either trapped by the reductase
CC T3R to form vindorosine and vindoline precursors, or it spontaneously
CC rearranges to form a member of the eburnamine class of monoterpene
CC indole alkaloids. {ECO:0000269|PubMed:25850027}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; JN613016; AEX07771.1; -; mRNA.
DR EMBL; KP122967; AKM12282.1; -; mRNA.
DR EMBL; LN831957; CQR79399.1; -; mRNA.
DR AlphaFoldDB; I1TEM1; -.
DR SMR; I1TEM1; -.
DR KEGG; ag:AKM12282; -.
DR BioCyc; MetaCyc:MON-20195; -.
DR BRENDA; 1.14.14.50; 1211.
DR UniPathway; UPA00365; -.
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: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:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
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 Alkaloid metabolism; Endoplasmic reticulum; Heme; Iron; Membrane;
KW Metal-binding; Monooxygenase; Oxidoreductase; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..506
FT /note="Tabersonine 3-oxygenase"
FT /id="PRO_0000439949"
FT TOPO_DOM 1..5
FT /note="Lumenal"
FT /evidence="ECO:0000305"
FT TRANSMEM 6..26
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 27..506
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT BINDING 450
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:Q96242"
SQ SEQUENCE 506 AA; 57443 MW; 2A19115E1F30B785 CRC64;
MEFHESSPFV FITRGFIFIA ISIAVLRRII SKKTKTLPPG PWKLPLIGNL HQFLGSVPYQ
ILRDLAQKNG PLMHLQLGEV SAIVAASPQM AKEITKTLDL QFADRPVIQA LRIVTYDYLD
ISFNAYGKYW RQLRKIFVQE LLTSKRVRSF CSIREDEFSN LVKTINSANG KSINLSKLMT
SCTNSIINKV AFGKVRYERE VFIDLINQIL ALAGGFKLVD LFPSYKILHV LEGTERKLWE
IRGKIDKILD KVIDEHRENS SRTGKGNGCN GQEDIVDILL RIEEGGDLDL DIPFGNNNIK
ALLFDIIAGG TETSSTAVDW AMSEMMRNPH VMSKAQKEIR EAFNGKEKIE ENDIQNLKYL
KLVIQETLRL HPPAPLLMRQ CREKCEIGGY HIPVGTKAFI NVWAIGRDPA YWPNPESFIP
ERFDDNTYEF TKSEHHAFEY LPFGAGRRMC PGISFGLANV ELPLALLLYH FNWQLPDGST
TLDMTEATGL AARRKYDLQL IATSYA
