C71Y1_CATRO
ID C71Y1_CATRO Reviewed; 516 AA.
AC W8JDE2; Q9AW94;
DT 13-FEB-2019, integrated into UniProtKB/Swiss-Prot.
DT 14-MAY-2014, sequence version 1.
DT 03-AUG-2022, entry version 22.
DE RecName: Full=Alstonine synthase {ECO:0000303|PubMed:29942076};
DE Short=CrAS {ECO:0000303|PubMed:29942076};
DE EC=1.14.14.- {ECO:0000269|PubMed:29942076};
DE AltName: Full=Cytochrome P450 71AY1 {ECO:0000303|PubMed:29942076};
DE Short=CrCYP71AY1 {ECO:0000303|PubMed:29942076};
GN Name=CYP71AY1 {ECO:0000303|PubMed:24710322};
GN Synonyms=CYP71 {ECO:0000303|PubMed:8507838};
GN ORFNames=Caros018961 {ECO:0000303|PubMed:24710322};
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;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=24710322; DOI=10.1038/ncomms4606;
RA Miettinen K., Dong L., Navrot N., Schneider T., Burlat V., Pollier J.,
RA Woittiez L., van der Krol S., Lugan R., Ilc T., Verpoorte R.,
RA Oksman-Caldentey K.M., Martinoia E., Bouwmeester H., Goossens A.,
RA Memelink J., Werck-Reichhart D.;
RT "The seco-iridoid pathway from Catharanthus roseus.";
RL Nat. Commun. 5:3606-3606(2014).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA] OF 6-516.
RC STRAIN=cv. G. Don;
RX PubMed=8507838; DOI=10.1007/bf00014944;
RA Meijer A.H., Souer E., Verpoorte R., Hoge J.H.C.;
RT "Isolation of cytochrome P-450 cDNA clones from the higher plant
RT Catharanthus roseus by a PCR strategy.";
RL Plant Mol. Biol. 22:379-383(1993).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, TISSUE
RP SPECIFICITY, AND MUTAGENESIS OF VAL-126; LYS-212; SER-220; VAL-308; GLU-310
RP AND VAL-452.
RX PubMed=29942076; DOI=10.1038/s41589-018-0078-4;
RA Dang T.T., Franke J., Carqueijeiro I.S.T., Langley C., Courdavault V.,
RA O'Connor S.E.;
RT "Sarpagan bridge enzyme has substrate-controlled cyclization and
RT aromatization modes.";
RL Nat. Chem. Biol. 14:760-763(2018).
CC -!- FUNCTION: Involved in monoterpene indole alkaloids (MIAs) biosynthesis
CC (PubMed:29942076). Converts by aromatization the tetrahydro-beta-
CC carboline alkaloids tetrahydroalstonine and ajmalicine to the
CC corresponding beta-carboline alkaloids alstonine and serpentine,
CC respectively (PubMed:29942076). {ECO:0000269|PubMed:29942076}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=A + O2 + reduced [NADPH--hemoprotein reductase] +
CC tetrahydroalstonine = AH2 + alstonine + H(+) + 2 H2O + oxidized
CC [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:58128, Rhea:RHEA-
CC COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:13193,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17499, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC ChEBI:CHEBI:142526, ChEBI:CHEBI:142530;
CC Evidence={ECO:0000269|PubMed:29942076};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=A + ajmalicine + O2 + reduced [NADPH--hemoprotein reductase] =
CC AH2 + H(+) + 2 H2O + oxidized [NADPH--hemoprotein reductase] +
CC serpentine; Xref=Rhea:RHEA:58132, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC COMP:11965, ChEBI:CHEBI:13193, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17499, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210, ChEBI:CHEBI:142527, ChEBI:CHEBI:142531;
CC Evidence={ECO:0000269|PubMed:29942076};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q96242};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=19.6 uM for tetrahydroalstonine {ECO:0000269|PubMed:29942076};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000250|UniProtKB:P0DO13}; Single-pass type II membrane protein
CC {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Highly expressed in stems (PubMed:29942076).
CC Expressed at low levels in roots (PubMed:29942076).
CC {ECO:0000269|PubMed:29942076}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
CC -!- WEB RESOURCE: Name=ORCAE database;
CC URL="https://bioinformatics.psb.ugent.be/orcae/overview/Catro";
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DR EMBL; KF309243; AHK60849.1; -; mRNA.
DR EMBL; AJ295719; CAC27827.1; -; mRNA.
DR AlphaFoldDB; W8JDE2; -.
DR SMR; W8JDE2; -.
DR KEGG; ag:AHK60849; -.
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:0016712; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen; IDA:UniProtKB.
DR GO; GO:0009709; P:terpenoid indole alkaloid biosynthetic process; IDA: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; Monooxygenase;
KW Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..516
FT /note="Alstonine synthase"
FT /id="PRO_0000446225"
FT TRANSMEM 6..26
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 453
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"
FT MUTAGEN 126
FT /note="V->A: No effect on activity towards
FT tetrahydroalstonine."
FT /evidence="ECO:0000269|PubMed:29942076"
FT MUTAGEN 212
FT /note="K->R: Reduces activity towards tetrahydroalstonine."
FT /evidence="ECO:0000269|PubMed:29942076"
FT MUTAGEN 220
FT /note="S->A: Acquires the capacity to convert
FT geissoschizine to polyneuridine aldehyde; no effect on
FT activity towards tetrahydroalstonine."
FT /evidence="ECO:0000269|PubMed:29942076"
FT MUTAGEN 308
FT /note="V->I: Reduces activity towards tetrahydroalstonine."
FT /evidence="ECO:0000269|PubMed:29942076"
FT MUTAGEN 310
FT /note="E->W: Acquires the capacity to convert
FT geissoschizine to polyneuridine aldehyde; slightly
FT increases activity towards tetrahydroalstonine."
FT /evidence="ECO:0000269|PubMed:29942076"
FT MUTAGEN 452
FT /note="V->I: Acquires the capacity to convert
FT geissoschizine to polyneuridine aldehyde; slightly
FT increases activity towards tetrahydroalstonine."
FT /evidence="ECO:0000269|PubMed:29942076"
SQ SEQUENCE 516 AA; 57776 MW; 0ADB7AC2692ADCD1 CRC64;
MDQLMNFSLT SPIFLLLSSL FLIILLNKLM RGNKIQKGKK LPPGPKKIAI IGNLPSNGRF
TSLIVFLNNL AEKYGPIMHL RIGQLSAVII SSAEKAKEIL NTHGVRVADR PQTTVAKIML
YNSLGVTFAP YGDYLKQLRQ IYAMELLSPK TVKSFWTIMD DELSTMITSI KSEVGQPMIL
HDKMMTYLYA MLCRATVGSV CNGRETLIMA AKETSALSAS IRIEDLFPSV KILPVISGLK
SKLTNLLKEL DIVLEDIISA REKKLLSQPQ QPLMLDEEDM LGVLLKYKNG KGNDTKFRVT
NNDIKAIVFE LILAGTLSSA AIVEWCMSEL MKNPELLKKA QDEVRQVLKG KKTISGSDVG
KLEYVKMVVK ESVRLHPPAP LLFPRECREE FEIDGMTIPK KSWVIINYWA IGRDPKIWPN
ADKFEPERFS NNNIDFYGSN FELIPFGAGR RVCPGILFGT TNVELLLAAF LFHFDWELPG
GMKPEELDMN ELFGAGCIRE NPLCLIPSIS TVVEGN
