C82X1_PAPSO
ID C82X1_PAPSO Reviewed; 540 AA.
AC I3V6B7;
DT 03-JUL-2019, integrated into UniProtKB/Swiss-Prot.
DT 05-SEP-2012, sequence version 1.
DT 03-AUG-2022, entry version 31.
DE RecName: Full=(13S,14R)-13-O-acetyl-1-hydroxy-N-methylcanadine 8-hydroxylase CYP82X1 {ECO:0000305};
DE EC=1.14.14.167 {ECO:0000269|PubMed:25485687, ECO:0000269|PubMed:27378283};
DE AltName: Full=Cytochrome P450 82X1 {ECO:0000303|PubMed:22653730};
GN Name=CYP82X1 {ECO:0000303|PubMed:22653730};
OS Papaver somniferum (Opium poppy).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Ranunculales; Papaveraceae; Papaveroideae;
OC Papaver.
OX NCBI_TaxID=3469;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=22527754; DOI=10.1007/s11103-012-9913-2;
RA Desgagne-Penix I., Farrow S.C., Cram D., Nowak J., Facchini P.J.;
RT "Integration of deep transcript and targeted metabolite profiles for eight
RT cultivars of opium poppy.";
RL Plant Mol. Biol. 79:295-313(2012).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND TISSUE SPECIFICITY.
RX PubMed=22653730; DOI=10.1126/science.1220757;
RA Winzer T., Gazda V., He Z., Kaminski F., Kern M., Larson T.R., Li Y.,
RA Meade F., Teodor R., Vaistij F.E., Walker C., Bowser T.A., Graham I.A.;
RT "A Papaver somniferum 10-gene cluster for synthesis of the anticancer
RT alkaloid noscapine.";
RL Science 336:1704-1708(2012).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=25485687; DOI=10.1038/nchembio.1717;
RA Dang T.T., Chen X., Facchini P.J.;
RT "Acetylation serves as a protective group in noscapine biosynthesis in
RT opium poppy.";
RL Nat. Chem. Biol. 11:104-106(2015).
RN [4]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=27378283; DOI=10.1038/ncomms12137;
RA Li Y., Smolke C.D.;
RT "Engineering biosynthesis of the anticancer alkaloid noscapine in yeast.";
RL Nat. Commun. 7:12137-12137(2016).
RN [5]
RP FUNCTION.
RX PubMed=29610307; DOI=10.1073/pnas.1721469115;
RA Li Y., Li S., Thodey K., Trenchard I., Cravens A., Smolke C.D.;
RT "Complete biosynthesis of noscapine and halogenated alkaloids in yeast.";
RL Proc. Natl. Acad. Sci. U.S.A. 115:E3922-E3931(2018).
CC -!- FUNCTION: Cytochrome P450 involved in the biosynthesis of the
CC benzylisoquinoline alkaloid noscapine (PubMed:25485687,
CC PubMed:27378283, PubMed:29610307). Converts (13S,14R)-13-O-acetyl-1-
CC hydroxy-N-methylcanadine to (13S,14R)-13-O-acetyl-1,8-dihydroxy-N-
CC methylcanadine (PubMed:25485687, PubMed:27378283).
CC {ECO:0000269|PubMed:25485687, ECO:0000269|PubMed:27378283,
CC ECO:0000269|PubMed:29610307}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(13S,14R)-13-O-acetyl-1-hydroxy-N-methylcanadine + O2 +
CC reduced [NADPH--hemoprotein reductase] = (13S,14R)-13-O-acetyl-1,8-
CC dihydroxy-N-methylcanadine + H(+) + H2O + oxidized [NADPH--
CC hemoprotein reductase]; Xref=Rhea:RHEA:57388, Rhea:RHEA-COMP:11964,
CC Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC ChEBI:CHEBI:141640, ChEBI:CHEBI:141642; EC=1.14.14.167;
CC Evidence={ECO:0000269|PubMed:25485687, ECO:0000269|PubMed:27378283};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:57389;
CC Evidence={ECO:0000305|PubMed:25485687};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q96242};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=4.9 uM for (13S,14R)-13-O-acetyl-1-hydroxy-N-methylcanadine
CC {ECO:0000269|PubMed:25485687};
CC Vmax=54.5 nmol/min/mg enzyme with (13S,14R)-13-O-acetyl-1-hydroxy-N-
CC methylcanadine as substrate {ECO:0000269|PubMed:25485687};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Highly expressed in capsules (PubMed:22653730).
CC Expressed is stems (PubMed:22653730). {ECO:0000269|PubMed:22653730}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; JN185333; AFK73719.1; -; mRNA.
DR EMBL; JQ659002; AFB74614.1; -; Genomic_DNA.
DR AlphaFoldDB; I3V6B7; -.
DR SMR; I3V6B7; -.
DR EnsemblPlants; RZC84729; RZC84729; C5167_047516.
DR Gramene; RZC84729; RZC84729; C5167_047516.
DR KEGG; ag:AFB74614; -.
DR OMA; WASEVIH; -.
DR BioCyc; MetaCyc:MON-20625; -.
DR BRENDA; 1.14.14.167; 4515.
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:0033075; P:isoquinoline alkaloid biosynthetic process; IEA:UniProt.
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; Heme; Iron; Membrane; Metal-binding; Monooxygenase;
KW Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..540
FT /note="(13S,14R)-13-O-acetyl-1-hydroxy-N-methylcanadine 8-
FT hydroxylase CYP82X1"
FT /id="PRO_0000447596"
FT TRANSMEM 15..35
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 483
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 540 AA; 61207 MW; C7AFB82F0F6F2A04 CRC64;
MELFIKLPFI QPIPFSIILV TTVSIVLLYS VFFWVTDKKK KRKKAPNAAG AWPLIGHLRL
LMNDKEPLYR ALGSMADKYG PAFNIRLGNQ EVLVVSNWEM VKQCFGNQND KLFSNRQTTL
AAKYMLNQTT SSGFAPYGPY WRELRKIMVQ QLLSKQSLES WKHLKIKEMD ASFSKLNELC
NNNGTGTATL IRMDEWFAEL TFNVIARNVF GYQSGGRSTA LTNGDTESKG ERYKKTLEEA
LHLMSIFAVS DIFPSLEWVD RLRGLIRNMK RFGDELNSIA GCLIEEHRQK RLQSVSKSDK
GVGDEQDFVD VLLSVAEKSQ LPGDDPDLVI KSMILEIVSG GSETTSSTLT WALCLLLNHP
HVLKKAKEEL DTHVGKDRHV EESDTPKLVY INAIIKESMR LYPNGAMLDR LALEECEVGG
FHVPAGGRLF VNVWKIQRDP SVWENPLEFK PERWFLSNGE KMDVDYKGHN HEFIPFGIGR
RMCAGMLWAS EVIHLVLPRL IHGFDMKAAS ANGKVDMAEM AGMVICFKKT PLEVMVNPRE
