NMT_CATRO
ID NMT_CATRO Reviewed; 289 AA.
AC W5U2K2; E3UTU0;
DT 10-MAY-2017, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 26.
DE RecName: Full=3-hydroxy-16-methoxy-2,3-dihydrotabersonine N-methyltransferase {ECO:0000305};
DE EC=2.1.1.99 {ECO:0000269|PubMed:20956330, ECO:0000269|PubMed:21802100};
DE AltName: Full=16-methoxy-2,3-dihydro-3-hydroxytabersonine N-methyltransferase {ECO:0000305};
DE AltName: Full=2,3-dihydrotabersonine N-methyltransferase {ECO:0000303|PubMed:20956330};
DE Short=CrDhtNMT {ECO:0000303|PubMed:20956330};
GN Name=NMT {ECO:0000303|PubMed:20956330};
GN Synonyms=Cr2270 {ECO:0000303|PubMed:20956330};
OS Catharanthus roseus (Madagascar periwinkle) (Vinca rosea).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Gentianales; Apocynaceae; Rauvolfioideae; Vinceae;
OC Catharanthinae; Catharanthus.
OX NCBI_TaxID=4058 {ECO:0000312|EMBL:AHH33092.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, SUBSTRATE SPECIFICITY, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, INDUCTION BY METHYL JASMONATE, AND
RP ACTIVITY REGULATION.
RX PubMed=20956330; DOI=10.1073/pnas.1009003107;
RA Liscombe D.K., Usera A.R., O'Connor S.E.;
RT "Homolog of tocopherol C methyltransferases catalyzes N methylation in
RT anticancer alkaloid biosynthesis.";
RL Proc. Natl. Acad. Sci. U.S.A. 107:18793-18798(2010).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], SUBSTRATE SPECIFICITY, AND SUBUNIT.
RX PubMed=26848097; DOI=10.1104/pp.15.01813;
RA Levac D., Cazares P., Yu F., De Luca V.;
RT "A picrinine N-methyltransferase belongs to a new family of gamma-
RT tocopherol-like methyltransferases found in medicinal plants that make
RT biologically active monoterpenoid indole alkaloids.";
RL Plant Physiol. 170:1935-1944(2016).
RN [3]
RP SUBCELLULAR LOCATION.
RX PubMed=16665811; DOI=10.1104/pp.85.4.1099;
RA De Luca V., Cutler A.J.;
RT "Subcellular localization of enzymes involved in indole alkaloid
RT biosynthesis in Catharanthus roseus.";
RL Plant Physiol. 85:1099-1102(1987).
RN [4]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=21802100; DOI=10.1016/j.phytochem.2011.07.001;
RA Liscombe D.K., O'Connor S.E.;
RT "A virus-induced gene silencing approach to understanding alkaloid
RT metabolism in Catharanthus roseus.";
RL Phytochemistry 72:1969-1977(2011).
CC -!- FUNCTION: S-adenosyl-L-methionine-dependent N-methyltransferase that
CC catalyzes a nitrogen methylation involved in vindoline biosynthesis
CC (PubMed:20956330, PubMed:21802100). Displays a strict requirement for a
CC 2,3-dihydro bond in the aspidosperma skeleton (PubMed:20956330). Can
CC use 2,3-dihydrotabersonine, 2,3-dihydro-3-hydroxytabersonine and
CC 2,3,6,7-tetraydro-3-hydroxytabersonine as substrates, but not
CC tabersonine, vincadifformine, picrinine, 21-hydroxycyclolochnericine,
CC tryptamine, norharmane, harmaline, catharanthine, norajmaline,
CC ajmaline, serpentine, ajmalicine, yohimbine or gamma-tocopherol
CC (PubMed:20956330, PubMed:26848097). {ECO:0000269|PubMed:20956330,
CC ECO:0000269|PubMed:21802100, ECO:0000269|PubMed:26848097}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(3R)-3-hydroxy-16-methoxy-2,3-dihydrotabersonine + S-adenosyl-
CC L-methionine = deacetoxyvindoline + H(+) + S-adenosyl-L-homocysteine;
CC Xref=Rhea:RHEA:11336, ChEBI:CHEBI:15378, ChEBI:CHEBI:57856,
CC ChEBI:CHEBI:57965, ChEBI:CHEBI:58485, ChEBI:CHEBI:59789; EC=2.1.1.99;
CC Evidence={ECO:0000269|PubMed:20956330, ECO:0000269|PubMed:21802100};
CC -!- ACTIVITY REGULATION: Inhibited by gamma-tocopherol.
CC {ECO:0000269|PubMed:20956330}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=8.8 uM for 2,3-dihydrotabersonine {ECO:0000269|PubMed:20956330};
CC KM=22.0 uM for S-adenosyl-L-methionine {ECO:0000269|PubMed:20956330};
CC Vmax=67.0 pmol/sec/mg enzyme toward 2,3-dihydrotabersonine
CC {ECO:0000269|PubMed:20956330};
CC Vmax=151.6 pmol/sec/mg enzyme toward S-adenosyl-L-methionine
CC {ECO:0000269|PubMed:20956330};
CC Note=kcat is 2.47 sec(-1) for 2,3-dihydrotabersonine. kcat is 5.4
CC sec(-1) for S-adenosyl-L-methionine. {ECO:0000269|PubMed:20956330};
CC -!- PATHWAY: Alkaloid biosynthesis; vindoline biosynthesis.
CC -!- SUBUNIT: Homodimer. {ECO:0000305|PubMed:26848097}.
CC -!- SUBCELLULAR LOCATION: Thylakoid {ECO:0000269|PubMed:16665811}.
CC -!- INDUCTION: Up-regulated upon methyl jasmonate treatment.
CC {ECO:0000269|PubMed:20956330}.
CC -!- SIMILARITY: Belongs to the class I-like SAM-binding methyltransferase
CC superfamily. gTMT family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=ADP00410.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR EMBL; HM584929; ADP00410.1; ALT_INIT; mRNA.
DR EMBL; KF896244; AHH33092.1; -; mRNA.
DR AlphaFoldDB; W5U2K2; -.
DR SMR; W5U2K2; -.
DR KEGG; ag:ADP00410; -.
DR UniPathway; UPA00365; -.
DR GO; GO:0009579; C:thylakoid; IEA:UniProtKB-SubCell.
DR GO; GO:0030768; F:16-methoxy-2,3-dihydro-3-hydroxytabersonine N-methyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0008170; F:N-methyltransferase activity; IDA:CACAO.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.150; -; 1.
DR InterPro; IPR013216; Methyltransf_11.
DR InterPro; IPR025774; MTs_g-TMT.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR Pfam; PF08241; Methyltransf_11; 1.
DR SUPFAM; SSF53335; SSF53335; 1.
DR PROSITE; PS51581; SAM_GTMT; 1.
PE 1: Evidence at protein level;
KW Alkaloid metabolism; Methyltransferase; S-adenosyl-L-methionine; Thylakoid;
KW Transferase.
FT CHAIN 1..289
FT /note="3-hydroxy-16-methoxy-2,3-dihydrotabersonine N-
FT methyltransferase"
FT /id="PRO_0000439951"
FT REGION 71..80
FT /note="SAM motif I"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00914"
FT REGION 134..142
FT /note="SAM motif II"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00914"
FT REGION 161..170
FT /note="SAM motif III"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00914"
FT CONFLICT 146
FT /note="E -> K (in Ref. 1; ADP00410)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 289 AA; 31789 MW; 17FE5F2FF5D6BE3D CRC64;
MEEKQEKVAE FYDKVTGAWD LFYGVHLHDG YYEPGTTATM AISQDAVIRM IDELLRFAGV
SEDPAKKPRS MLDVGSGLGG TCVYVAKKYD IQCTGITISP NQVKYAQDYA ATEGVENKVS
FDCGDALDMP YSDGKFDVVF TINCIEHVHD KEKFIREMVR VAAPGAAIII ASQAHPNLSP
GESLKPRDKK ILQKICDGAG AVSLCSSDDY VRWLTPLPVK EIKAADWTQN ITPLYPLLMK
EAFTWKGFTS IVLKGGWRAI NLINAVRLVA KAANDGILKF AVVTGRKSI
