Information on EC 2.1.1.53 - putrescine N-methyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.53
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RECOMMENDED NAME
GeneOntology No.
putrescine N-methyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + putrescine = S-adenosyl-L-homocysteine + N-methylputrescine
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
methyl group transfer
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
N-methyl-Delta;1-pyrrolinium cation biosynthesis
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Tropane, piperidine and pyridine alkaloid biosynthesis
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Metabolic pathways
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:putrescine N-methyltransferase
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CAS REGISTRY NUMBER
COMMENTARY hide
9075-39-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + 1,4-diamino-2,3-dibromo-trans-but-2-ene
S-adenosyl-L-homocysteine + ?
show the reaction diagram
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-
-
-
?
S-adenosyl-L-methionine + 1,4-diamino-2-hydroxybutane
S-adenosyl-L-homocysteine + ?
show the reaction diagram
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-
-
-
?
S-adenosyl-L-methionine + 1,4-diamino-2-oxobutane
S-adenosyl-L-homocysteine + ?
show the reaction diagram
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-
-
-
?
S-adenosyl-L-methionine + 1,4-diamino-trans-but-2-ene
S-adenosyl-L-homocysteine + ?
show the reaction diagram
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-
-
-
?
S-adenosyl-L-methionine + 4-(aminomethyl)piperidine
S-adenosyl-L-homocysteine + ?
show the reaction diagram
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-
-
-
?
S-adenosyl-L-methionine + N-methylputrescine
S-adenosylhomocysteine + N,N-dimethylputrescine
show the reaction diagram
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about 8% of activity compared to putrescine
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-
?
S-adenosyl-L-methionine + putrescine
S-adenosyl-L-homocysteine + N-methylputrescine
show the reaction diagram
S-adenosyl-L-methionine + putrescine
S-adenosylhomocysteine + N-methylputrescine
show the reaction diagram
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-
-
-
?
S-adenosyl-L-methionine + trans-1,4-diamonocyclohexane
S-adenosyl-L-homocysteine + ?
show the reaction diagram
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-
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + putrescine
S-adenosyl-L-homocysteine + N-methylputrescine
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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no requirement of cofactors
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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Mg2+, Mn2+ and EDTA at 0.001 M have no influence
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,3-Diaminopropane
1,6-diaminohexane
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weaker inhibitor
1,7-Diaminoheptane
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weaker inhibitor
2-(aminomethyl)pyridine
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3-methylcadaverine
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4-(aminomethyl)pyridine
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Ca2+
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28% inhibition by 5 mM
cadaverine
Cr2+
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31% inhibition by 5 mM
Cu2+
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73% inhibition by 5 mM
EDTA
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23% inhibition by 5 mM
Fe2+
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33% inhibition by 5 mM
Fe3+
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14% inhibition by 5 mM
Mg2+
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28% inhibition by 5 mM
Mn2+
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26% inhibition by 5 mM
n-butylamine
N-methyl-1,3-diaminopropane
N-methylputrescine
Ni2+
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71% inhibition by 5 mM
p-chloromercuribenzoate
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strong
S-adenosyl-L-homocysteine
Zn2+
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27% inhibition by 5 mM
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cysteine
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slight activatory effect
methyl jasmonate
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0758 - 3.13
putrescine
0.013 - 0.15
S-adenosyl-L-methionine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.047 - 5.14
putrescine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04
cadaverine
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0.15
N-methylputrescine
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0.01
S-adenosyl-L-homocysteine
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00016
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leaves
0.0018
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roots
0.051
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putrescine
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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pI: 5.3
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26
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assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
isoform PMT1, calculated from amino acid sequence
6.3
isoform PMT2, calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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acropetal and basipetal regions
Manually annotated by BRENDA team
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apical cells, increases with age
Manually annotated by BRENDA team
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transcript only in potato tuber eyes at sprouting start and in 1 and 3 mm long sprouts
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37220
isoform PMT1, calculated from amino acid sequence
38120
isoform PMT2, calculated from amino acid sequence
40000
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gel filtration
60000
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gel filtration
360000
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SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 37000, SDS-PAGE
homodimer
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gel filtration
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
homology-based modeling of Datura stramonium spermidine synthase SPDS1 and putrescine N-methyltransferase PMT
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9.5
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20% loss of activity
485445
9.8
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35% loss of activity
485445
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol stabilizes
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.05 M phosphate buffer, pH 7.5, 0.01 M 2-mercaptoethanol, 12.5% glucose, stable for several weeks
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-20C, 12.5% glucose for at least a week
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0C, without addition of a sulfhydryl compound, most activity is lost within a few days
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4C, glycine-NaOH buffer, pH 9, stable for 1 month
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by HiTrap affinity chromatography and gel filtration
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by Ni2+ chelate affinity chromatography
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by Ni2+-chelate affinity chromatography
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gel filtration, approximately 95% pure
gel filtration, approximately 95% pure; gel filtration, approximately 95% pure
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AbPMT1 is expressed more strongly than ABPMT2 in the root
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AbPMT1 is expressed more strongly than ABPMT2 in the root; AbPMT1 is expressed more strongly than ABPMT2 in the root
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expressed in Escherichia coli
expressed in Escherichia coli M15[pREP4] cells
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expressed in Escherichia coli Rosetta-gami (DE3), transformed with a pET-21d vector encoding pmt
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expressed in Escherichia coli, three genes isolated from Nicotiana sylvestris: NsPMT1, NsPMT2, NsPMT3; expressed in Escherichia coli, three genes isolated from Nicotiana sylvestris: NsPMT1, NsPMT2, NsPMT3; expressed in Escherichia coli, three genes isolated from Nicotiana sylvestris: NsPMT1, NsPMT2, NsPMT3
expressed in Escherichia coli; expressed in Escherichia coli
expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli
expressed in roots and leaves
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expressed in Rosetta-gami (DE3) cells; expressed in Rosetta-gami (DE3) cells; into vector pET21d and transferred into Escherichia coli Rosetta-gami (DE3) cells; into vector pET21d and transferred into Escherichia coli Rosetta-gami (DE3) cells
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expressed in Rosetta-gami (DE3) cells; into vector pET21d and transferred into Escherichia coli Rosetta-gami (DE3) cells
expressed in xylem-tissues of roots, three genes isolated from Nicotiana sylvestris: NsPMT1, NsPMT2, NsPMT3, a highly conserved NsPMT 5'-flanking region is sufficient for tissue-specific and jasmonate-responsive expression of all three PMT genes; expressed in xylem-tissues of roots, three genes isolated from Nicotiana sylvestris: NsPMT1, NsPMT2, NsPMT3, a highly conserved NsPMT 5'-flanking region is sufficient for tissue-specific and jasmonate-responsive expression of all three PMT genes; expressed in xylem-tissues of roots, three genes isolated from Nicotiana sylvestris: NsPMT1, NsPMT2, NsPMT3, a highly conserved NsPMT 5'-flanking region is sufficient for tissue-specific and jasmonate-responsive expression of all three PMT genes
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into pETat-32a(+) vector and expressed in Escherichia coli BL21trxB(DE3)
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pmt1 and pmt2 cloned into pET-21d vector, PMT1 expressed in Escherichia coli Rosetta-gami (DE3), PMT2 expressed in Escherichia coli BL21 (DE3) and Rosetta-gami (DE3), pmt1 yields active enzymes, while pmt2 expression results in insoluble protein
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tobacco pmt gene with a selectable marker (nptII) gene and the T-DNA of pRiA4 of Agrobacterium rhizogenes introduced into Hyoscyamus niger leaf explants using disarmed Agrobacterium tumefaciens C58C1 strain
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
Ag+ increases expression of putrescine N-methyltransferase I (maximum after 48 h). The expression of PMT1 reaches the highest level at 24 h treatment in ethanol
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dark conditions and the absence of auxin upregulate enzyme promoter activity, with light being dominant to the repressive effects of auxin. Under highly repressive conditions for alkaloid synthesis, including normal culture conditions in the light, both reactive oxygen species scavengers result in significant induction of enzyme promoter activity. Treatment of callus with catalase results in the upregulation of enzyme promoter activity
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isoform PMT1 expression is upregulated by methyl jasmonate in all tissues, reaching the highest level after 24 h of the treatment; isoform PMT2 expression is upregulated by methyl jasmonate in all tissues, reaching the highest level after 24 h of the treatment
treatments with diphenyleneiodinium chloride, Rose Bengal diacetate, and 1,2-dihydroxybenzene-3,5-disulfonic acid do not induce or repress enzyme expression
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under normal callus growth conditions in the presence of light and auxin, enzyme promoter activity is strongly repressed
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H108L
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40% maximal turn over velocity of wild-type, Km for S-adenosyl-L-methionine is slightly elevated, but putrescine saturation appears only at very high concentrations
I136V
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reduced PMT activity, about half of the wild-type activity
I141D
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mutant shows no activity; no PMT activity
T117Q
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mutant shows no activity; no PMT activity
T117Q/I141D
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mutant shows no activity; no PMT activity
additional information
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cDNA from Nicotiana tabacum encoding PMT is introduced into the genome of a scopolamine-producing Hyoscyamus niger mediated by the disarmed Agrobacterium tumefaciens strain C58C1, which also carries Agrobacterium rhizogenes Ri plasmid pRiA4, and expressed under the control of the CaMV 35S promoter. Hairy root lines transformed with pmt present 5fold higher PMT activity than the control, and the methylputrescine levels of the resulting engineered hairy roots increase up to 5fold compared to the control and wild-type roots, but no increase in tropane alkaloids measured. After methyl jasmonate treatment, a considerable increase of PMTase and endogenous H6Hase as well as an increase in scopolamine content is found either in the transgenic hairy roots or the control
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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convenient and accurate enzyme-coupled colorimetric PMT assay, based on the conversion of S-adenosyl-L-homocysteine by 5'-methylthioacenosine/S-adenosylhomocysteine nucleosidase and S-ribosylhomocysteine lyase
synthesis
additional information