C82E4_NICTO
ID C82E4_NICTO Reviewed; 517 AA.
AC A1YJE3;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 06-FEB-2007, sequence version 1.
DT 03-AUG-2022, entry version 53.
DE RecName: Full=Nicotine N-demethylase CYP82E4 {ECO:0000303|PubMed:17102129};
DE Short=NND {ECO:0000303|PubMed:17102129};
DE EC=1.14.14.- {ECO:0000269|PubMed:17102129};
DE AltName: Full=Cytochrome P450 82E4 {ECO:0000303|PubMed:17102129};
DE Short=NtomCYP82E4 {ECO:0000303|PubMed:17102129};
GN Name=CYP82E4 {ECO:0000303|PubMed:17102129};
OS Nicotiana tomentosiformis (Tobacco).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Solanales; Solanaceae; Nicotianoideae; Nicotianeae;
OC Nicotiana.
OX NCBI_TaxID=4098;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, PATHWAY,
RP BIOPHYSICOCHEMICAL PROPERTIES, TISSUE SPECIFICITY, AND DEVELOPMENTAL STAGE.
RX PubMed=17102129; DOI=10.1074/jbc.m609512200;
RA Gavilano L.B., Coleman N.P., Bowen S.W., Siminszky B.;
RT "Functional analysis of nicotine demethylase genes reveals insights into
RT the evolution of modern tobacco.";
RL J. Biol. Chem. 282:249-256(2007).
CC -!- FUNCTION: Involved in the biosynthesis of pyridine alkaloid natural
CC products, leading mainly to the production of anabasine, anatabine,
CC nicotine and nornicotine, effective deterrents against herbivores with
CC antiparasitic and pesticide properties (neurotoxins); nornicotine
CC serves as the precursor in the synthesis of the carcinogen compound N'-
CC nitrosonornicotine (NNN) (PubMed:17102129). Catalyzes the demethylation
CC of nicotine to form nornicotine (PubMed:17102129).
CC {ECO:0000269|PubMed:17102129}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-nicotine + O2 + reduced [NADPH--hemoprotein reductase] =
CC (S)-nornicotine + formaldehyde + H(+) + H2O + oxidized [NADPH--
CC hemoprotein reductase]; Xref=Rhea:RHEA:70999, Rhea:RHEA-COMP:11964,
CC Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16842, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210, ChEBI:CHEBI:59806, ChEBI:CHEBI:190184;
CC Evidence={ECO:0000269|PubMed:17102129};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:71000;
CC Evidence={ECO:0000269|PubMed:17102129};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:P04798};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=9.1 uM for nicotine {ECO:0000269|PubMed:17102129};
CC Vmax=1.0 nmol/min/mg enzyme with nicotine as substrate
CC {ECO:0000269|PubMed:17102129};
CC -!- PATHWAY: Alkaloid biosynthesis; nicotine biosynthesis.
CC {ECO:0000269|PubMed:17102129}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Expressed in leaves. {ECO:0000269|PubMed:17102129}.
CC -!- DEVELOPMENTAL STAGE: Expressed in green leaves but fades out during
CC senescence. {ECO:0000269|PubMed:17102129}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. CYP82E2 subfamily.
CC {ECO:0000305}.
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DR EMBL; EF042307; ABM46920.1; -; Genomic_DNA.
DR RefSeq; XP_009609539.1; XM_009611244.2.
DR GeneID; 104103347; -.
DR KEGG; nto:104103347; -.
DR OrthoDB; 702827at2759; -.
DR BRENDA; 1.14.14.B11; 3648.
DR UniPathway; UPA00107; -.
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 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; Isopeptide bond; Membrane; Metal-binding;
KW Monooxygenase; Oxidoreductase; Transmembrane; Transmembrane helix;
KW Ubl conjugation.
FT CHAIN 1..517
FT /note="Nicotine N-demethylase CYP82E4"
FT /id="PRO_0000455782"
FT TRANSMEM 2..22
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 457
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:P04798"
FT CROSSLNK 254
FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with
FT G-Cter in ubiquitin)"
FT /evidence="ECO:0000250|UniProtKB:Q9SZU1"
SQ SEQUENCE 517 AA; 59328 MW; CF099EEB9AEFB91A CRC64;
MLSPIEAIVG LVTFTFLFYF LWTKKSQKPS KPLPPKIPGG WPVIGHLFHF NDDGDDRPLA
RKLGDLADKY GPVFTFRLGL PLVLVVSSYE AVKDCFSTND AIFSNRPAFL YGDYLGYNNA
MLFLANYGPY WRKNRKLVIQ EVLSASRLEK FKHVRFARIQ ASIKNLYTRI DGNSSTINLT
DWLEELNFGL IVKMIAGKNY ESGKGDEQVE RFKKAFKDFM ILSMEFVLWD AFPIPLFKWV
DFQGHVKAMK RTFKDIDSVF QNWLEEHINK REKMEVNAEG NEQDFIDVVL SKMSNEYLGE
GYSRDTVIKA TVFSLVLDAA DTVALHINWG MALLINNQKA LTKAQEEIDT KVGKDRWVEE
SDIKDLVYLQ AIVKEVLRLY PPGPLLVPHE NVEDCVVSGY HIPKGTRLFA NVMKLQRDPK
LWSDPDTFDP ERFIATDIDF RGQYYKYIPF GSGRRSCPGM TYALQVEHLT MAHLIQGFNY
RTPNDEPLDM KEGAGITIRK VNPVELIIAP RLAPELY
