C96T1_NARAP
ID C96T1_NARAP Reviewed; 513 AA.
AC A0A140IL90;
DT 10-MAY-2017, integrated into UniProtKB/Swiss-Prot.
DT 11-MAY-2016, sequence version 1.
DT 03-AUG-2022, entry version 19.
DE RecName: Full=Noroxomaritidine synthase {ECO:0000303|PubMed:26941773};
DE EC=1.14.19.50 {ECO:0000269|PubMed:26941773};
DE AltName: Full=CYP96T1 {ECO:0000303|PubMed:26941773};
DE AltName: Full=Cytochrome P450 96T1 {ECO:0000303|PubMed:26941773};
GN Name=Cyp96T1 {ECO:0000303|PubMed:26941773};
OS Narcissus aff. pseudonarcissus MK-2014 (Daffodil).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Liliopsida; Asparagales; Amaryllidaceae;
OC Amaryllidoideae; Narcissus.
OX NCBI_TaxID=1540222;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBSTRATE SPECIFICITY, AND PATHWAY.
RX PubMed=26941773; DOI=10.3389/fpls.2016.00225;
RA Kilgore M.B., Augustin M.M., May G.D., Crow J.A., Kutchan T.M.;
RT "CYP96T1 of Narcissus sp. aff. pseudonarcissus catalyzes formation of the
RT para-para' C-C phenol couple in the Amaryllidaceae alkaloids.";
RL Front. Plant Sci. 7:225-225(2016).
CC -!- FUNCTION: Cytochrome P450 that catalyzes an intramolecular para-para'
CC C-C phenol coupling of 4'-O-methylnorbelladine in alkaloids
CC biosynthesis, including haemanthamine- and crinamine-type alkaloids,
CC promising anticancer agents. Catalyzes the formation of (10bR,4aS)-
CC noroxomaritidine and (10bS,4aR)-noroxomaritidine from 4'-O-
CC methylnorbelladine. Produces also N-demethylnarwedine as a minor
CC product. Involved in the biosynthesis of haemanthamine. Can use also
CC 4'-O-methyl-N-methylnorbelladine, (S)- and (R)-coclaurine as
CC substrates, but not 3'-O-methylnorbelladine, 3',4'-O-
CC dimethylnorbelladine, norbelladine, haemanthamine, (10bS,4aR)- or
CC (10bR,4aS)-noroxomaritidine, isovanillin or tyramine.
CC {ECO:0000269|PubMed:26941773}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4'-O-methylnorbelladine + O2 + reduced [NADPH--hemoprotein
CC reductase] = (10bS,4aR)-noroxomaritidine + H(+) + 2 H2O + oxidized
CC [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:51264, 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:133993, ChEBI:CHEBI:133996;
CC EC=1.14.19.50; Evidence={ECO:0000269|PubMed:26941773};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4'-O-methylnorbelladine + O2 + reduced [NADPH--hemoprotein
CC reductase] = (10bR,4aS)-noroxomaritidine + H(+) + 2 H2O + oxidized
CC [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:51260, 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:133993, ChEBI:CHEBI:133995;
CC EC=1.14.19.50; Evidence={ECO:0000269|PubMed:26941773};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000305};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.13 uM for 4'-O-methylnorbelladine {ECO:0000269|PubMed:26941773};
CC KM=3.28 uM for 4'-O-methyl-N-methylnorbelladine
CC {ECO:0000269|PubMed:26941773};
CC KM=637 uM for (S)-coclaurine {ECO:0000269|PubMed:26941773};
CC KM=659 uM for (R)-coclaurine {ECO:0000269|PubMed:26941773};
CC Note=kcat is 15.0 min(-1) with 4'-O-methylnorbelladine as substrate.
CC kcat is 2.44 min(-1) with 4'-O-methyl-N-methylnorbelladine as
CC substrate. kcat is 1.34 min(-1) with (S)-coclaurine as substrate.
CC kcat is 2.07 min(-1) with (R)-coclaurine as substrate.
CC {ECO:0000269|PubMed:26941773};
CC pH dependence:
CC Optimum pH is 6.5. {ECO:0000269|PubMed:26941773};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius.
CC {ECO:0000269|PubMed:26941773};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:26941773}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; KT693311; AMO65741.1; -; mRNA.
DR AlphaFoldDB; A0A140IL90; -.
DR SMR; A0A140IL90; -.
DR KEGG; ag:AMO65741; -.
DR BioCyc; MetaCyc:MON-20066; -.
DR BRENDA; 1.14.19.50; 14577.
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 GO; GO:0009820; P:alkaloid metabolic process; 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; Membrane; Metal-binding; Monooxygenase;
KW Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..513
FT /note="Noroxomaritidine synthase"
FT /id="PRO_0000439852"
FT TRANSMEM 14..34
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:Q96242"
SQ SEQUENCE 513 AA; 59001 MW; 6310A26C0A7E1A81 CRC64;
MATSSSAWLM FSDHYPEILI AIACFLIFSL LLSARSSSED SLPYNWPIFG MLPAIISNNQ
FNDFTTARLR KMGWTFIFKG PWLLDMDYIF TCDPSNINHM FNDNFENYPK GELGKVFDIF
GNNIFNADGD LWHDHRKMAQ TILWDGNYRT MQATFIRNKM DNALIPILDS AASKRKPVDL
QDVFFRFTFD TSCFSVLAAD PESLTMEFPA VPFSKAADQA LDAALTRHIT PRLIWKLKRF
FNVGSERTLA VAWKVIDSYI YDKIAELKAK RKLVGKINSY DAVSFYMDNF NIHDDKFLRD
NAFTYLLAQR NTQSLTMTWL FYALFENPKV ELKILSELKS IVDESSERKF NDGFALFDSN
MIQSAIYLHA TLCEALRIYP PVPFEIKDAH KADVLPSGHK VRAGEKILFS PYAMARMKGI
WGDDCLEFKP ERWITGNGTL KHEPAYKFFA FSAGPRICLG KELSFTQMKM VVATIIYNFH
LQMVKGHVVE QSNSILMDMK HGLMVQVRKR SVM
