C82E4_TOBAC
ID C82E4_TOBAC Reviewed; 517 AA.
AC Q38Q87; L7Y094; Q078P0; Q078P1; Q078P2; Q078P3; Q078P4; Q078P5; Q078P6;
AC Q078P7; Q078P8; Q38Q86;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 03-AUG-2022, sequence version 2.
DT 03-AUG-2022, entry version 81.
DE RecName: Full=Nicotine N-demethylase CYP82E4 {ECO:0000303|Ref.3};
DE Short=NND {ECO:0000303|Ref.3};
DE EC=1.14.14.- {ECO:0000269|PubMed:16192354, ECO:0000269|PubMed:17102129, ECO:0000269|Ref.3};
DE AltName: Full=Cytochrome P450 82E4 {ECO:0000303|PubMed:16192354, ECO:0000303|Ref.2, ECO:0000303|Ref.3, ECO:0000303|Ref.4};
DE Short=NtCYP82E4 {ECO:0000303|PubMed:16192354, ECO:0000303|Ref.2, ECO:0000303|Ref.3, ECO:0000303|Ref.4};
DE Short=NtabCYP82E4 {ECO:0000303|PubMed:17102129};
GN Name=CYP82E4 {ECO:0000303|PubMed:16192354, ECO:0000303|Ref.2,
GN ECO:0000303|Ref.3, ECO:0000303|Ref.4};
GN Synonyms=CYP82E4v1 {ECO:0000303|PubMed:16192354, ECO:0000303|Ref.2};
GN ORFNames=LOC107819427 {ECO:0000312|RefSeq:XP_016501019.1};
OS Nicotiana tabacum (Common 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=4097;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], VARIANT 2-VAL-PHE-3 DELINS LEU-SER, FUNCTION,
RP CATALYTIC ACTIVITY, AND PATHWAY.
RC STRAIN=cv. Burley DH91-1307-46, and cv. Petit Havana;
RX PubMed=16192354; DOI=10.1073/pnas.0506581102;
RA Siminszky B., Gavilano L., Bowen S.W., Dewey R.E.;
RT "Conversion of nicotine to nornicotine in Nicotiana tabacum is mediated by
RT CYP82E4, a cytochrome P450 monooxygenase.";
RL Proc. Natl. Acad. Sci. U.S.A. 102:14919-14924(2005).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], AND VARIANTS GLN-61; LEU-74; ASP-99; VAL-191;
RP GLU-217; SER-231; ALA-252; GLU-309; GLY-314; MET-335; CYS-353; VAL-410;
RP VAL-413; LEU-416; PRO-423; GLY-439; PRO-452; ILE-471 AND SER-488.
RC STRAIN=cv. Burley 4407;
RA Xu D., Shen Y., Nielsen M.T.;
RT "Cloning of cytochrome P450 genes from Nicotiana.";
RL Submitted (SEP-2005) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, PATHWAY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=cv. Burley 4407;
RX DOI=10.1111/j.1399-3054.2006.00811.x;
RA Xu D., Shen Y., Chappell J., Cui M., Nielsen M.T.;
RT "Biochemical and molecular characterizations of nicotine demethylase in
RT tobacco.";
RL Physiol. Plantarum 129:307-319(2007).
RN [4]
RP NUCLEOTIDE SEQUENCE [MRNA].
RC STRAIN=cv. B37;
RA Yang J., Sun B., Zhang F., Zhou G.-J.;
RT "Cloning of CYP82E4 gene in Nicotiana tabacum.";
RL Submitted (NOV-2012) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. TN90;
RX PubMed=24807620; DOI=10.1038/ncomms4833;
RA Sierro N., Battey J.N., Ouadi S., Bakaher N., Bovet L., Willig A.,
RA Goepfert S., Peitsch M.C., Ivanov N.V.;
RT "The tobacco genome sequence and its comparison with those of tomato and
RT potato.";
RL Nat. Commun. 5:3833-3833(2014).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, PATHWAY, DEVELOPMENTAL STAGE, AND TISSUE
RP SPECIFICITY.
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).
RN [7]
RP REVIEW ON ALKALOID BIOSYNTHESIS IN NICOTIANA TABACUM.
RX PubMed=23953973; DOI=10.1016/j.phytochem.2013.06.002;
RA Dewey R.E., Xie J.;
RT "Molecular genetics of alkaloid biosynthesis in Nicotiana tabacum.";
RL Phytochemistry 94:10-27(2013).
RN [8]
RP REVIEW ON NICOTINE BIOSYNTHESIS.
RX PubMed=25582664; DOI=10.1007/s00438-015-0989-7;
RA Wang X., Bennetzen J.L.;
RT "Current status and prospects for the study of Nicotiana genomics,
RT genetics, and nicotine biosynthesis genes.";
RL Mol. Genet. Genomics 290:11-21(2015).
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) (Ref.3, PubMed:17102129, PubMed:16192354).
CC Catalyzes the demethylation of nicotine to form nornicotine (Ref.3,
CC PubMed:17102129, PubMed:16192354). {ECO:0000269|PubMed:16192354,
CC ECO:0000269|PubMed:17102129, ECO:0000269|Ref.3}.
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:16192354, ECO:0000269|PubMed:17102129,
CC ECO:0000269|Ref.3};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:71000;
CC Evidence={ECO:0000269|PubMed:16192354, ECO:0000269|PubMed:17102129,
CC ECO:0000269|Ref.3};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:P04798};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.9 uM for nicotine {ECO:0000269|Ref.3};
CC Vmax=8.9 pmol/sec/mg enzyme with nicotine as substrate
CC {ECO:0000269|Ref.3};
CC -!- PATHWAY: Alkaloid biosynthesis; nicotine biosynthesis.
CC {ECO:0000269|PubMed:16192354, ECO:0000269|PubMed:17102129,
CC ECO:0000269|Ref.3}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Expressed at low levels in green leaves.
CC {ECO:0000269|PubMed:17102129}.
CC -!- DEVELOPMENTAL STAGE: Highly induced during senescence.
CC {ECO:0000269|PubMed:17102129}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. CYP82E2 subfamily.
CC {ECO:0000305}.
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DR EMBL; DQ131885; ABA07804.1; -; mRNA.
DR EMBL; DQ131886; ABA07805.1; -; mRNA.
DR EMBL; DQ219341; ABD39474.1; -; mRNA.
DR EMBL; DQ219342; ABD39475.1; -; mRNA.
DR EMBL; DQ219343; ABD39476.1; -; mRNA.
DR EMBL; DQ219344; ABD39477.1; -; mRNA.
DR EMBL; DQ219345; ABD39478.1; -; mRNA.
DR EMBL; DQ219346; ABD39479.1; -; mRNA.
DR EMBL; DQ219347; ABD39480.1; -; mRNA.
DR EMBL; DQ219348; ABD39481.1; -; mRNA.
DR EMBL; DQ219349; ABD39482.1; -; mRNA.
DR EMBL; DQ205656; ABB36475.1; -; mRNA.
DR EMBL; KC120817; AGD93125.1; -; mRNA.
DR EMBL; EF370115; ABN42695.1; -; Genomic_DNA.
DR RefSeq; NP_001312976.1; NM_001326047.1.
DR RefSeq; XP_016501019.1; XM_016645533.1.
DR SMR; Q38Q87; -.
DR GeneID; 107819427; -.
DR KEGG; nta:107819427; -.
DR OMA; MREFTHA; -.
DR OrthoDB; 702827at2759; -.
DR BioCyc; MetaCyc:MON-12449; -.
DR BRENDA; 1.14.14.1; 3645.
DR BRENDA; 1.14.14.B11; 3645.
DR UniPathway; UPA00107; -.
DR Proteomes; UP000084051; Unplaced.
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:0008168; F:methyltransferase activity; IEA:UniProtKB-KW.
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:0098542; P:defense response to other organism; IBA:GO_Central.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
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 Methyltransferase; Monooxygenase; Oxidoreductase; Reference proteome;
KW Transferase; Transmembrane; Transmembrane helix; Ubl conjugation.
FT CHAIN 1..517
FT /note="Nicotine N-demethylase CYP82E4"
FT /id="PRO_0000455781"
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"
FT VARIANT 2..3
FT /note="VF -> LS (in CYP82E4v2)"
FT /evidence="ECO:0000269|PubMed:16192354"
FT VARIANT 61
FT /note="R -> Q (in CYP82E4v11)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 74
FT /note="F -> L (in CYP82E4v4)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 99
FT /note="N -> D (in CYP82E4v11)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 191
FT /note="I -> V (in CYP82E4v7)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 217
FT /note="K -> E (in CYP82E4v7)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 231
FT /note="A -> S (in CYP82E4v10)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 252
FT /note="T -> A (in CYP82E4v8)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 309
FT /note="K -> E (in CYP82E4v12)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 314
FT /note="S -> G (in CYP82E4v11)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 335
FT /note="I -> M (in CYP82E4v10)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 353
FT /note="G -> C (in CYP82E4v12)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 410
FT /note="A -> V (in CYP82E4v8)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 413
FT /note="M -> V (in CYP82E4v5)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 416
FT /note="Q -> L (in CYP82E4v6)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 423
FT /note="S -> P (in CYP82E4v6)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 439
FT /note="D -> G (in CYP82E4v10)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 452
FT /note="S -> P (in CYP82E4v12)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 471
FT /note="M -> I (in CYP82E4v11)"
FT /evidence="ECO:0000269|Ref.2"
FT VARIANT 488
FT /note="L -> S (in CYP82E4v3)"
FT /evidence="ECO:0000269|Ref.2"
FT CONFLICT 417
FT /note="R -> C (in Ref. 4; AGD93125)"
FT /evidence="ECO:0000305"
FT CONFLICT 436
FT /note="T -> I (in Ref. 4; AGD93125)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 517 AA; 59359 MW; 7F099DA8A9A9456B CRC64;
MVFPIEAIVG LVTFTFLFFF 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
