PNMT_THLFG
ID PNMT_THLFG Reviewed; 356 AA.
AC C3SBW0;
DT 27-JUL-2011, integrated into UniProtKB/Swiss-Prot.
DT 16-JUN-2009, sequence version 1.
DT 03-AUG-2022, entry version 40.
DE RecName: Full=Pavine N-methyltransferase {ECO:0000303|PubMed:19624470};
DE Short=PfPavNMT {ECO:0000303|PubMed:19624470};
DE EC=2.1.1.300 {ECO:0000269|PubMed:19624470, ECO:0000269|PubMed:27573242};
OS Thalictrum flavum subsp. glaucum (Yellow meadow rue).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Ranunculales; Ranunculaceae; Thalictroideae;
OC Thalictrum.
OX NCBI_TaxID=150095;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND INDUCTION.
RX PubMed=19624470; DOI=10.1111/j.1365-313x.2009.03980.x;
RG Natural Products Genomics Resource (NAPGEN);
RA Liscombe D.K., Ziegler J., Schmidt J., Ammer C., Facchini P.J.;
RT "Targeted metabolite and transcript profiling for elucidating enzyme
RT function: isolation of novel N-methyltransferases from three
RT benzylisoquinoline alkaloid-producing species.";
RL Plant J. 60:729-743(2009).
RN [2] {ECO:0007744|PDB:5KN4, ECO:0007744|PDB:5KOC, ECO:0007744|PDB:5KOK, ECO:0007744|PDB:5KPC, ECO:0007744|PDB:5KPG}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) IN COMPLEX WITH
RP S-ADENOSYL-L-METHIONINE; S-ADENOSYL-L-HOMOCYSTEINE;
RP (R)-TETRAHYDROPAPAVERINE AND (S)-TETRAHYDROPAPAVERINE, CATALYTIC ACTIVITY,
RP SUBSTRATE SPECIFICITY, SUBUNIT, AND MUTAGENESIS OF TYR-79; GLU-80; GLU-205
RP AND HIS-206.
RX PubMed=27573242; DOI=10.1074/jbc.m116.747261;
RA Torres M.A., Hoffarth E., Eugenio L., Savtchouk J., Chen X., Morris J.S.,
RA Facchini P.J., Ng K.K.;
RT "Structural and functional studies of pavine N-methyltransferase from
RT Thalictrum flavum reveal novel insights into substrate recognition and
RT catalytic mechanism.";
RL J. Biol. Chem. 291:23403-23415(2016).
CC -!- FUNCTION: N-methyltransferase with a substrate preference for (+-)-
CC pavine, but also active with (R,S)-stylopine, (R,S)-scoulerine, (R,S)-
CC tetrahydropalmatine, (R,S)-tetrahydropapaverine and (S)-reticuline
CC (PubMed:19624470, PubMed:27573242). No activity with (R)-reticuline
CC (PubMed:27573242). In the presence of a racemic mixture of
CC tetrahydropapaverine (THP), one molecule of (S)-THP binds in a
CC productive mode, while one molecule of (R)-THP is bound next to it in a
CC non-productive mode (PubMed:27573242). The (R)-THP may inhibit the
CC release of products from the enzyme when higher concentrations of the
CC racemic substrate are added to the reaction (PubMed:27573242).
CC {ECO:0000269|PubMed:19624470, ECO:0000269|PubMed:27573242}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(+-)-pavine + S-adenosyl-L-methionine = H(+) + N-methylpavine
CC + S-adenosyl-L-homocysteine; Xref=Rhea:RHEA:39979, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:76921,
CC ChEBI:CHEBI:76922; EC=2.1.1.300;
CC Evidence={ECO:0000269|PubMed:19624470, ECO:0000269|PubMed:27573242};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=47 uM for (+-)-pavine {ECO:0000269|PubMed:19624470};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:27573242}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:19624470}.
CC -!- INDUCTION: Not induced by elicitor treatment.
CC {ECO:0000269|PubMed:19624470}.
CC -!- SIMILARITY: Belongs to the CFA/CMAS family. {ECO:0000305}.
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DR EMBL; EU883010; ACO90251.1; -; mRNA.
DR PDB; 5KN4; X-ray; 1.99 A; A/B=1-356.
DR PDB; 5KOC; X-ray; 2.29 A; A/B=1-356.
DR PDB; 5KOK; X-ray; 1.79 A; A/B=1-356.
DR PDB; 5KPC; X-ray; 2.50 A; A/B=1-356.
DR PDB; 5KPG; X-ray; 1.60 A; A/B=1-356.
DR PDBsum; 5KN4; -.
DR PDBsum; 5KOC; -.
DR PDBsum; 5KOK; -.
DR PDBsum; 5KPC; -.
DR PDBsum; 5KPG; -.
DR AlphaFoldDB; C3SBW0; -.
DR SMR; C3SBW0; -.
DR KEGG; ag:ACO90251; -.
DR BRENDA; 2.1.1.300; 9013.
DR SABIO-RK; C3SBW0; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0008168; F:methyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.150; -; 1.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR SUPFAM; SSF53335; SSF53335; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Methyltransferase; S-adenosyl-L-methionine;
KW Transferase.
FT CHAIN 1..356
FT /note="Pavine N-methyltransferase"
FT /id="PRO_0000411114"
FT ACT_SITE 331
FT /evidence="ECO:0000250"
FT BINDING 96..97
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:27573242,
FT ECO:0007744|PDB:5KOC, ECO:0007744|PDB:5KPC"
FT BINDING 135..140
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:27573242,
FT ECO:0007744|PDB:5KOC"
FT BINDING 159..163
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:27573242,
FT ECO:0007744|PDB:5KOC"
FT BINDING 185..186
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:27573242,
FT ECO:0007744|PDB:5KOC, ECO:0007744|PDB:5KPC"
FT BINDING 201
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:27573242,
FT ECO:0007744|PDB:5KOC"
FT BINDING 205
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:27573242,
FT ECO:0007744|PDB:5KOK"
FT MUTAGEN 79
FT /note="Y->A: Loss of catalytic activity with (S)-reticuline
FT and racemic pavine, but increased catalytic activity with
FT racemic tetrahydropapaverine."
FT /evidence="ECO:0000269|PubMed:27573242"
FT MUTAGEN 80
FT /note="E->A: Increased catalytic activity with (S)-
FT reticuline, racemic pavine and racemic
FT tetrahydropapaverine."
FT /evidence="ECO:0000269|PubMed:27573242"
FT MUTAGEN 205
FT /note="E->A: Strongly decreased catalytic activity. Over
FT 90% decreased catalytic activity; when associated with A-
FT 206."
FT /evidence="ECO:0000269|PubMed:27573242"
FT MUTAGEN 206
FT /note="H->A: Strongly decreased catalytic activity. Over
FT 90% decreased catalytic activity; when associated with A-
FT 205."
FT /evidence="ECO:0000269|PubMed:27573242"
FT HELIX 11..19
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 25..43
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 48..62
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 73..78
FT /evidence="ECO:0007829|PDB:5KOK"
FT HELIX 83..89
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 102..105
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 108..122
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 130..134
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 140..148
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 153..159
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 161..173
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 178..183
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 186..188
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 195..202
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 204..206
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 210..218
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 221..236
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 244..246
FT /evidence="ECO:0007829|PDB:5KPC"
FT HELIX 250..254
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 261..264
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 267..270
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 273..283
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 286..301
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 303..314
FT /evidence="ECO:0007829|PDB:5KPG"
FT HELIX 317..343
FT /evidence="ECO:0007829|PDB:5KPG"
FT STRAND 345..355
FT /evidence="ECO:0007829|PDB:5KPG"
SQ SEQUENCE 356 AA; 41556 MW; D4DA5BE5C7192B00 CRC64;
METKQTKKEA VANLIKRIEH GEVSDEEIRG MMKIQVQKRL KWGYKPTHEQ QLAQLVTFAQ
SLKGMEMAEE VDTLDAELYE IPLPFLHIMC GKTLKFSPGY FKDESTTLDE SEVYMMDLYC
ERAQIKDGQS ILDLGCGHGS LTLHVAQKYR GCKVTGITNS VSQKEFIMDQ CKKLDLSNVE
IILEDVTKFE TEITYDRIFA VALIEHMKNY ELFLKKVSTW IAQYGLLFVE HHCHKVFAYQ
YEPLDEDDWY TEYIFPSGTL VMSSSSILLY FQEDVSVVNH WTLSGKHPSL GFKQWLKRLD
DNIDEVKEIF ESFYGSKEKA MKFITYWRVF CIAHSQMYST NNGEEWMLSQ VLFKKK
