ID   P6H_ESCCA               Reviewed;         524 AA.
AC   F2Z9C1;
DT   03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT   28-JUN-2011, sequence version 1.
DT   03-AUG-2022, entry version 35.
DE   RecName: Full=Protopine 6-monooxygenase;
DE            EC=1.14.14.98 {ECO:0000269|PubMed:22421633};
DE   AltName: Full=Protopine 6-hydroxylase;
DE            Short=P6H;
GN   Name=CYP82N2v2;
OS   Eschscholzia californica (California poppy).
OC   Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC   Spermatophyta; Magnoliopsida; Ranunculales; Papaveraceae;
OC   Eschscholzioideae; Eschscholzia.
OX   NCBI_TaxID=3467;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP   BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX   PubMed=22421633; DOI=10.1016/j.phytochem.2012.02.013;
RA   Takemura T., Ikezawa N., Iwasa K., Sato F.;
RT   "Molecular cloning and characterization of a cytochrome P450 in
RT   sanguinarine biosynthesis from Eschscholzia californica cells.";
RL   Phytochemistry 91:100-108(2013).
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: Catalyzes the conversion of protopine and allocryptopine to
CC       dihydrosanguinarine and dihydrochelerythrine, respectively, in the
CC       biosynthesis of isoquinoline alkaloid sanguinarine.
CC       {ECO:0000269|PubMed:22421633}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=O2 + protopine + reduced [NADPH--hemoprotein reductase] = 6-
CC         hydroxyprotopine + H(+) + H2O + oxidized [NADPH--hemoprotein
CC         reductase]; Xref=Rhea:RHEA:22644, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC         COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC         ChEBI:CHEBI:16415, ChEBI:CHEBI:17104, ChEBI:CHEBI:57618,
CC         ChEBI:CHEBI:58210; EC=1.14.14.98;
CC         Evidence={ECO:0000269|PubMed:22421633};
CC   -!- COFACTOR:
CC       Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=6.45 uM for protopine {ECO:0000269|PubMed:22421633};
CC         KM=8.70 uM for allocryptopine {ECO:0000269|PubMed:22421633};
CC   -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:22421633}.
CC   -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane {ECO:0000305};
CC       Multi-pass membrane protein {ECO:0000305}.
CC   -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR   EMBL; AB598834; BAK20464.1; -; mRNA.
DR   KEGG; ag:BAK20464; -.
DR   BioCyc; MetaCyc:MON-12344; -.
DR   BRENDA; 1.14.14.98; 2173.
DR   SABIO-RK; F2Z9C1; -.
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:0047087; F:protopine 6-monooxygenase activity; IDA:UniProtKB.
DR   GO; GO:0033075; P:isoquinoline 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..524
FT                   /note="Protopine 6-monooxygenase"
FT                   /id="PRO_0000430258"
FT   TRANSMEM        4..24
FT                   /note="Helical; Name=1"
FT                   /evidence="ECO:0000255"
FT   TRANSMEM        232..252
FT                   /note="Helical; Name=2"
FT                   /evidence="ECO:0000255"
FT   TRANSMEM        319..339
FT                   /note="Helical; Name=3"
FT                   /evidence="ECO:0000255"
FT   BINDING         462
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"
SQ   SEQUENCE   524 AA;  59462 MW;  F8167484A2299237 CRC64;
     MDSLMLAYLF PISVASIIAF VFLYNLFSSR TLKNKKIRTA PMATGAWPVL GHLHLFGSGE
     LPHKMLAAMA DKYGSAFRMK FGKHTTLVVS DTRIVKECFT TNDTLFSNRP STKAFQLMTY
     DNESVAFTPY GSYWREIRKI STLKLLSNHR LQAIKDVRAS EVNVCFKTLY DQCKNPSGSA
     PILIDMKKWF EEVSNNVVMR VIVGRQNFGS KIVQGEEEAI HYKKVMDELL RLASLSMFSD
     FAPLLGFVDI FQGNLSAMKR NAKKVDAILE NWLEEHRKKK NSVAESQQDF MDVMLSIVEE
     SKLSGHDADA VIKATCLAMI MGGTDTTAVS LTWIISLLMN NRHALKKARE ELDALVGKDR
     QVEDSDLKNL VYMNAIVKET MRMYPLGTLL ERETKEDCEI DGFHVKGGTR LLVNVWKLQR
     DPNVWVDPTE FRPERFLTEN ADIDVGGQHF ELLPFGAGRR VCPGVXFALQ FMHLVLARLI
     HGYDLNTLNE ENVDLTESPE GHVNHKASPL DLILTPRLHY KLYE