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Information on EC 2.1.1.300 - pavine N-methyltransferase
for references in articles please use BRENDA:EC2.1.1.300
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EC Tree 2 Transferases
2.1 Transferring one-carbon groups
2.1.1 Methyltransferases
2.1.1.300 pavine N-methyltransferase
IUBMB Comments
The enzyme, isolated from the plant Thalictrum flavum, also methylates (R,S)-stylopine and (S)-scoulerine (11%) with lower activity (14% and 11%, respectively).
The enzyme appears in viruses and cellular organisms
- 2.1.1.300
- catecholamine
-
sympathetic
-
end-stage
-
monoamine
-
flavoprotein
-
tone
- epinephrine
-
dinucleotide-dependent
-
catecholamine-metabolizing
-
hemodialyzed
Reaction Schemes
showSynonyms
pavine n-methyltransferase, pavine nmt, pavnmt, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + (+/-)-pavine = S-adenosyl-L-homocysteine + N-methylpavine
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-
-
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SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:(+/-)-pavine N-methyltransferase
The enzyme, isolated from the plant Thalictrum flavum, also methylates (R,S)-stylopine and (S)-scoulerine (11%) with lower activity (14% and 11%, respectively).
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + (+/-)-N-methylpavine
S-adenosyl-L-homocysteine + (+/-)-N,N-dimethylpavine
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (+/-)-pavine
S-adenosyl-L-homocysteine + (+/-)-N-methylpavine
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (+/-)-pavine
S-adenosyl-L-homocysteine + N-methylpavine
-
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (R,S)-scoulerine
S-adenosyl-L-homocysteine + N-methylscoulerine
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (S)-reticuline
S-adenosyl-L-homocysteine + (S)-tetrahydropapaverine
Substrates: best substrate. (R)-reticuline is not accepted as a substrate, revealing a strong stereoselectivity for (S)-reticuline
Products: -
Products: -
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S-adenosyl-L-methionine + (S)-scoulerine
S-adenosyl-L-homocysteine + N-methyl-(S)-scoulerine
-
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (S)-tetrahydropapaverine
S-adenosyl-L-homocysteine + (S)-laudanosine
additional information
?
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Substrates: analysis of substrate specificity by mass spectrometry, overview. Enzyme PavNMT exhibits a preference for (+)-pavine and the 1-benzylisoquinoline (S)-reticuline, but also accepts the protoberberines scoulerine and stylopine and, to a lesser extent, tetrahydropapaverine (THP). No activity with (R)-reticuline and papaverine, as well as cryptopine, glaucine, codeine, canadaline, noscapine, and berbamine. Structural determinants of substrate binding are derived from crystal structure analysis
Products: -
Products: -
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S-adenosyl-L-methionine + (R,S)-stylopine
S-adenosyl-L-homocysteine + N-methylstylopine
-
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (R,S)-stylopine
S-adenosyl-L-homocysteine + N-methylstylopine
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (S)-tetrahydropapaverine
S-adenosyl-L-homocysteine + (S)-laudanosine
Substrates: -
Products: -
Products: -
-
S-adenosyl-L-methionine + (S)-tetrahydropapaverine
S-adenosyl-L-homocysteine + (S)-laudanosine
Substrates: i.e. THP, low activity, a racemic mixture of THP is converted by PavNMT to laudanosine. The benzyl group of (S)-THP is most deeply buried in the substrate-binding pocket, whereas the isoquinoline moiety lies in a more exposed position where the plane of the six-carbon ring is stacked in a parallel arrangement against the benzyl group of a molecule of (R)-THP. The molecule of (R)-THP appears to be bound more loosely, with fewer direct contacts with the protein than seen for the molecule of (S)-THP. The loop formed by residues 75-91 covers both THP molecules, and the structure of this loop appears to be dependent on the binding of both THP molecules, because it is disordered when THP is not bound
Products: -
Products: -
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + (+/-)-N-methylpavine
S-adenosyl-L-homocysteine + (+/-)-N,N-dimethylpavine
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (+/-)-pavine
S-adenosyl-L-homocysteine + (+/-)-N-methylpavine
Substrates: -
Products: -
Products: -
?
S-adenosyl-L-methionine + (S)-tetrahydropapaverine
S-adenosyl-L-homocysteine + (S)-laudanosine
Substrates: -
Products: -
Products: -
-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
S-adenosyl-L-methionine
SAM, enzyme binding structure, overview
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(S)-tetrahydropapaverine
substrate inhibition at higher concentrations, enzyme complex structure with S-adenosyl-L-homocysteine
additional information
no substrate inhibition by (S)-reticuline
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.016
(S)-tetrahydropapaverine
pH 7.5, 30°C, recombinant enzyme
additional information
additional information
steady-state kinetics
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00079
(S)-tetrahydropapaverine
pH 7.5, 30°C, recombinant enzyme
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.12
(S)-tetrahydropapaverine
pH 7.5, 30°C, recombinant enzyme
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
the enzyme is involved in benzylisoquinoline alkaloids biosynthesis
additional information
substrate recognition and catalytic mechanism of pavine N-methyltransferase from Thalictrum flavum, highly conserved residues at the active site, overview. The loop formed by residues 75-91 covers both THP molecules, and the structure of this loop appears to be dependent on the binding of both THP molecules, because it is disordered when THP is not bound
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PNMT_THLFG
356
0
41556
Swiss-Prot
other Location (Reliability: 1)
A0A2P6PD90_ROSCH
147
0
16970
TrEMBL
other Location (Reliability: 3)
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
PavNMT forms a dimer with 2fold rotational symmetry in the asymmetric unit that is not seen in homologous NMTs
additional information
the overall structure of PavNMT reveals the presence of an alpha/beta SAM-binding domain with a canonical Rossmann fold that is shared among most SAM-dependent methyltransferases. In addition to this domain, a primarily alpha-helical BIA-binding domain forms the majority of the putative binding site for the methyl group acceptor
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
sitting-drop vapour-diffusion method, hanging-drop vapour-diffusion method at 16°C, crystallized in space group P2(1). The structure is solved at 2.0 A resolution using a xenon derivative and the single isomorphous replacement with anomalous scattering method
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purified recombinant native and selenomethionine-substituted wild-type and mutant H206A enzymes in complex with S-adenosyl-L-methionine, of wild-type apoenzyme, and of wild-type enzyme in complex with S-adenosyl-L-homocysteine (SAH) or with SAH and tetrahydropapaverine, hanging drop vapour diffusion method, mixing of 0.0015 ml of protein solution with 0.0015 ml of reservoir solution containing 340 g/l pentaerythritol ethoxylate (15:4 EO/OH), 50 mM bis-Tris-HCl, pH 6.0, 70 mM ammonium sulfate, and 160 g/l glycerol, and equilibration against 0.45 ml of reservoir solution at 22°C, 1-3 weeks, for the complex crystals, the enzyme is incubated with the ligands (0.75 mM THP oand/or 1 mM SAM/SAH) before crystallization, method optimization, X-ray diffraction structure determination and analysis at 1.6-2.3 A resolution
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E205A
site-directed mutagenesis, replacement of the residue with Ala results in a small decrease in protein stability as observed through a slight reduction in melting temperature, and to a large decrease in activity for three different substrates, especially the best substrate identified so far, (S)-reticuline
E205A/H206A
site-directed mutagenesis, replacement of Glu205 and His206 with Ala leads to large decreases in activity for three different substrates to about 10% of wild-type activity at pH 7.5
E80A
site-directed mutagenesis, the mutant shows 2fold increased activity compared to wild-type with substrate (S)-reticuline
H206A
site-directed mutagenesis, substrate binding structures compared to the wild-type enzyme, overview. Replacement of the residue with Ala results in a small decrease in protein stability as observed through a slight reduction in melting temperature, and to a large decrease in activity for three different substrates, especially the best substrate identified so far, (S)-reticuline
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged enzyme from Escherichia coli by nickel affinity chromatography, dialysis, anion exchange chromatography, and ultrafiltration
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His6-tagged wild-type and mutant enzymes and of selenomethionine-substituted enzyme in Escherichia coli
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Jain, A.; Ziegler, J.; Liscombe, D.K.; Facchini, P.J.; Tucker, P.A.; Panjikar, S.
Purification, crystallization and X-ray diffraction analysis of pavine N-methyltransferase from Thalictrum flavum
Acta Crystallogr. Sect. F
64
1066-1069
2008
Thalictrum flavum
brenda
Torres, M.A.; Hoffarth, E.; Eugenio, L.; Savtchouk, J.; Chen, X.; Morris, J.S.; Facchini, P.J.; Ng, K.K.
Structural and functional studies of pavine N-methyltransferase from Thalictrum flavum reveal novel insights into substrate recognition and catalytic mechanism
J. Biol. Chem.
291
23403-23415
2016
Thalictrum flavum subsp. glaucum (C3SBW0)
brenda
EXTERNAL LINKS (specific for EC-Number 2.1.1.300)
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Release 2023.1 (January 2023)
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