C82E3_NICTO
ID C82E3_NICTO Reviewed; 518 AA.
AC A1YJE2;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 06-FEB-2007, sequence version 1.
DT 03-AUG-2022, entry version 49.
DE RecName: Full=Nicotine N-demethylase CYP82E3 {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 82E3 {ECO:0000303|PubMed:17102129};
DE Short=NtomCYP82E3 {ECO:0000303|PubMed:17102129};
GN Name=CYP82E3 {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=48.1 uM for nicotine {ECO:0000269|PubMed:17102129};
CC Vmax=7.1 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: Highly expressed in green leaves but rapidly fades
CC out during senescence. {ECO:0000269|PubMed:17102129}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. CYP82E2 subfamily.
CC {ECO:0000305}.
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DR EMBL; EF042306; ABM46919.1; -; Genomic_DNA.
DR RefSeq; XP_009609537.1; XM_009611242.2.
DR GeneID; 104103346; -.
DR KEGG; nto:104103346; -.
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..518
FT /note="Nicotine N-demethylase CYP82E3"
FT /id="PRO_0000455784"
FT TRANSMEM 2..22
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 458
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 518 AA; 59601 MW; 1E35CA3C8E795258 CRC64;
MVFPVEAIVG LVTFTFLFYF LWTKKSQKPS KPLPPKIPGG WPVIGHLFYF DDDGDDRPLA
RKLGDLADKY GPVFTFRLGL PLVLVVSSYE AIKDCFSTND AIFSNRPAFL YGEYLGYNNA
MLFLANYGSY WRKNRKLIIQ EVLSASRLEK FKHVRFARIQ TSIKNLYTRI DGNSSTINLT
DWLEELNFGL IVKMIAGKNY ESGKGDEQVE RFKKAFKDFM ILSMEFVLWD AFPIPLFKWV
DFQGHVKAMK RTFKDIDSVF QNWLEEHIKK REKIMEVGTE GNEQDFIDVV LSKMSNEYLG
EGYSRDTVIK ATVFSLVLDA ADTVALHINW GMALLINNQN ALKKAQEEID TKVGKDRWVE
ESDIKDLVYL QAIVKEVLRL YPPGPLLVPH ENVEDCVVSG YHIPKGTRLF ANVMKLQRDP
KLWSNPDKFN PERFIARDID FHGQHYEYIP FGSGRRSCPG MTYALQVEHL TMAHLIQGFN
YRTPTDEPLD MKEGAGITIR KVNPVKVIIT PRLAPELY
