ID MSH_PAPSO Reviewed; 526 AA.
AC L7X3S1;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 03-APR-2013, sequence version 1.
DT 13-SEP-2023, entry version 33.
DE RecName: Full=Methyltetrahydroprotoberberine 14-monooxygenase {ECO:0000303|PubMed:23313486};
DE EC=1.14.14.97 {ECO:0000269|PubMed:23313486, ECO:0000269|Ref.2};
DE AltName: Full=(S)-cis-N-methylstylopine 14-hydroxylase {ECO:0000303|PubMed:23313486};
DE AltName: Full=(S)-cis-N-methyltetrahydroprotoberberine-14-hydroxylase {ECO:0000303|PubMed:23313486, ECO:0000303|Ref.2};
DE AltName: Full=Cytochrome P450 82N4 {ECO:0000303|PubMed:23313486};
DE AltName: Full=Methyltetrahydroprotoberberine 14-hydroxylase {ECO:0000303|PubMed:23313486};
DE AltName: Full=N-methylstylopine hydroxylase {ECO:0000303|PubMed:23313486};
DE Short=PsMSH {ECO:0000303|PubMed:23313486};
GN Name=CYP82N4 {ECO:0000303|PubMed:23313486};
GN and
GN Name=MSH {ECO:0000303|PubMed:23313486};
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], FUNCTION, PATHWAY, CATALYTIC ACTIVITY, TISSUE
RP SPECIFICITY, AND INDUCTION BY ELICITOR.
RX PubMed=23313486; DOI=10.1016/j.bbrc.2012.12.129;
RA Beaudoin G.A.W., Facchini P.J.;
RT "Isolation and characterization of a cDNA encoding (S)-cis-N-
RT methylstylopine 14-hydroxylase from opium poppy, a key enzyme in
RT sanguinarine biosynthesis.";
RL Biochem. Biophys. Res. Commun. 431:597-603(2013).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, PATHWAY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP ACTIVITY REGULATION.
RX DOI=10.1016/S0040-4039(00)96715-7;
RA Rueffer M., Zenk M.H.;
RT "Enzymatic formation of protopines by a microsomal cytochrome P-450 system
RT of Corydalis vaginans.";
RL Tetrahedron Lett. 28:5307-5310(1987).
CC -!- FUNCTION: Involved in the biosynthesis of the isoquinoline alkaloid
CC sanguinarine (PubMed:23313486, Ref.2). Catalyzes the conversion of N-
CC methylated protoberberine alkaloids N-methylstylopine and N-
CC methylcanadine into protopine and allocryptopine, respectively
CC (PubMed:23313486, Ref.2). Can also use (S)-cis-N-
CC methyltetrahydrothalifendine and (S)-cis-N-methyltetrahydropalmatine as
CC substrates (Ref.2). {ECO:0000269|PubMed:23313486, ECO:0000269|Ref.2}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-cis-N-methylcanadine + O2 + reduced [NADPH--hemoprotein
CC reductase] = allocryptopine + 2 H(+) + H2O + oxidized [NADPH--
CC hemoprotein reductase]; Xref=Rhea:RHEA:23684, Rhea:RHEA-COMP:11964,
CC Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17390, ChEBI:CHEBI:50540,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210; EC=1.14.14.97;
CC Evidence={ECO:0000269|PubMed:23313486, ECO:0000269|Ref.2};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-cis-N-methylstylopine + O2 + reduced [NADPH--hemoprotein
CC reductase] = 2 H(+) + H2O + oxidized [NADPH--hemoprotein reductase] +
CC protopine; Xref=Rhea:RHEA:76035, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC COMP:11965, ChEBI:CHEBI:444, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16415, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210; EC=1.14.14.97;
CC Evidence={ECO:0000269|PubMed:23313486, ECO:0000269|Ref.2};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-cis-N-methyltetrahydrothalifendine + O2 + reduced [NADPH--
CC hemoprotein reductase] = 7-hydroxy-8-methoxy-11-methyl-17,19-dioxa-
CC 11-azatetracyclo[12.7.0.0(4,9).0(16,20)]henicosa-
CC 1(21),4(9),5,7,14,16(20)-hexaen-2-one + 2 H(+) + H2O + oxidized
CC [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:76039, Rhea:RHEA-
CC COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210, ChEBI:CHEBI:194521, ChEBI:CHEBI:194522;
CC EC=1.14.14.97; Evidence={ECO:0000269|Ref.2};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-cis-N-methyltetrahydropalmatine + O2 + reduced [NADPH--
CC hemoprotein reductase] = 2 H(+) + H2O + muramine + oxidized [NADPH--
CC hemoprotein reductase]; Xref=Rhea:RHEA:76043, 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:194514, ChEBI:CHEBI:194523; EC=1.14.14.97;
CC Evidence={ECO:0000269|Ref.2};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q96242};
CC -!- ACTIVITY REGULATION: Repressed by cytochrome P450 inhibitors
CC ketoconazole, metyrapone, prochloraz, ancymidol and cytochrome C.
CC {ECO:0000269|Ref.2}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=12.5 uM for cis-N-methylcanadine {ECO:0000269|Ref.2};
CC KM=62.5 uM for NADPH {ECO:0000269|Ref.2};
CC pH dependence:
CC Optimum pH is 8.5. {ECO:0000269|Ref.2};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius. {ECO:0000269|Ref.2};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:23313486,
CC ECO:0000269|Ref.2}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Mainly expressed in roots, and barely in stems,
CC leaves and carpels. {ECO:0000269|PubMed:23313486}.
CC -!- INDUCTION: Induced by elicitor. {ECO:0000269|PubMed:23313486}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; KC154003; AGC92398.1; -; mRNA.
DR AlphaFoldDB; L7X3S1; -.
DR SMR; L7X3S1; -.
DR KEGG; ag:AGC92398; -.
DR BioCyc; MetaCyc:MONOMER-18686; -.
DR GO; GO:0016020; C:membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0047084; F:methyltetrahydroprotoberberine 14-monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0033075; P:isoquinoline alkaloid biosynthetic process; IDA:UniProtKB.
DR CDD; cd20654; CYP82; 1.
DR Gene3D; 1.10.630.10; Cytochrome P450; 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 PANTHER; PTHR47947; CYTOCHROME P450 82C3-RELATED; 1.
DR PANTHER; PTHR47947:SF19; CYTOCHROME P450 82C3-RELATED; 1.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; Cytochrome P450; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Heme; Iron; Membrane; Metal-binding; Monooxygenase; Oxidoreductase;
KW Transmembrane; Transmembrane helix.
FT CHAIN 1..526
FT /note="Methyltetrahydroprotoberberine 14-monooxygenase"
FT /id="PRO_0000430260"
FT TRANSMEM 14..34
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 468
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 526 AA; 59346 MW; 37F4A26D4AA15864 CRC64;
MRTESIKTNR PMDLLLQYLQ PISVALVVIA LVWNYGRRNP TKKLAPEASG GRPIMGHLHL
FNDGELTHRK LGAMADTYGP VFNIRFGSHK TLVVSDWEIV KECFTTNDKL FSNRPGTLGI
KLMFYDADSV GYAPYGAYWR DLRKISTLKL LSNHRIDTIK HLRSSEVESC FESLYSQWGN
GEKSGEFAPV RMDSWLGDLT FNVVARIVAG KKNFSANGDV GAQRYKAAMD EAMRLMRFFA
FSDVIPSLSW LDNLRGLVRE MKKCASEIDS IMATWVEEHR VKRNSGGNSQ LEHDFIDVCL
DIMEHSSLPG DDPDLVVKST CLDMILGGSD TTTVTLTWAM SLLLNHPQVL QKAKEELETQ
VGKNRQVDDS DIPNLPFIQA IIKETMRLYP AGPLIERRTM EDCEVAGYQV PAGTRLLVNV
WKMQRDGNVY KGDPLEFRPD RFLTSNADVD LKGQHYELIP FGAGRRICPG VSFAVQLMHL
VLARLLHEFE ITTVEPETKV DMAESGGLLC YKIMPLEVLI KPRLEI
//
