Information on EC 2.1.1.274 - salicylate 1-O-methyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.274
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RECOMMENDED NAME
GeneOntology No.
salicylate 1-O-methyltransferase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + salicylate = S-adenosyl-L-homocysteine + methyl salicylate
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
volatile benzenoid biosynthesis I (ester formation)
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SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:salicylate 1-O-methyltransferase
The enzyme, which is found in flowering plants, also has the activity of EC 2.1.1.273, benzoate O-methyltransferase.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
snapdragon
UniProt
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
Lillium hybrid cultivar
var. Yelloween, a popular OT (oriental x trumpet) hybrid, collected from the field in Xiaotangshan, Beijing, gene LiBSMT
UniProt
Manually annotated by BRENDA team
gene NbSAMT
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-
Manually annotated by BRENDA team
gene PbBSMT
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
Lillium hybrid cultivar
the enzyme belongs to SABATH family, a class of O-methyltransferases and N-methyltransferases
malfunction
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a knockout mutant fails to accumulate methyl salicylate following pathogen infection. These plants also fail to accumulate salicylate or its glucoside in the uninoculated leaves and do not develop systemic acquired resistance. However, the mutant exhibits normal levels of effector-triggered immunity and pathogen-associated molecular pattern-triggered immunity to Pseudomonas syringae and Hyaloperonospora arabidopsidis
metabolism
-
the enzyme is critical for methyl salicylate synthesis
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 + 3-hydroxybenzoate
S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate
show the reaction diagram
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1.8% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + 3-hydroxybenzoic acid
S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate
show the reaction diagram
S-adenosyl-L-methionine + 4-aminosalicylate
S-adenosyl-L-homocysteine + methyl 4-aminosalicylate
show the reaction diagram
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2% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + 4-hydroxybenzoate
S-adenosyl-L-homocysteine + methyl 4-hydroxybenzoate
show the reaction diagram
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lower catalytic efficiency with 4-hydroxybenzoate compared to salicylate
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-
?
S-adenosyl-L-methionine + anthranilate
?
show the reaction diagram
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9.3% activity compared to salicylate
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-
?
S-adenosyl-L-methionine + anthranilic acid
S-adenosyl-L-homocysteine + methyl anthranilate
show the reaction diagram
S-adenosyl-L-methionine + benzoate
?
show the reaction diagram
-
16.9% activity compared to salicylate
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-
?
S-adenosyl-L-methionine + benzoate
S-adenosyl-L-homocysteine + methyl benzoate
show the reaction diagram
-
2% activity compared to salicylate
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-
?
S-adenosyl-L-methionine + cinnamic acid
S-adenosyl-L-homocysteine + methyl cinnamate
show the reaction diagram
less than 2% relative activity at 1 mM methyl acceptor compared to activity with salicylate set at 100%
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-
?
S-adenosyl-L-methionine + jasmonic acid
S-adenosyl-L-homocysteine + methyl jasmonate
show the reaction diagram
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Y147S/M150H double mutant and Y147S/M150H/F347Y triple mutant are able to turn over jasmonic acid, while preserving substantial salicylate methylating activity
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?
S-adenosyl-L-methionine + salicylate
S-adenosyl-L-homocysteine + methyl salicylate
show the reaction diagram
S-adenosyl-L-methionine + salicylic acid
S-adenosyl-L-homocysteine + methyl salicylate
show the reaction diagram
S-adenosyl-L-methionine + vanillic acid
S-adenosyl-L-homocysteine + methyl 4-hydroxy-3-methoxybenzoate
show the reaction diagram
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5.1% activity of wild-type enzyme compared to salicylate methylation
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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 + 3-hydroxybenzoate
S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate
show the reaction diagram
-
1.8% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + 4-aminosalicylate
S-adenosyl-L-homocysteine + methyl 4-aminosalicylate
show the reaction diagram
-
2% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + 4-hydroxybenzoate
S-adenosyl-L-homocysteine + methyl 4-hydroxybenzoate
show the reaction diagram
-
lower catalytic efficiency with 4-hydroxybenzoate compared to salicylate
-
-
?
S-adenosyl-L-methionine + anthranilate
?
show the reaction diagram
-
9.3% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + benzoate
?
show the reaction diagram
-
16.9% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + benzoate
S-adenosyl-L-homocysteine + methyl benzoate
show the reaction diagram
-
2% activity compared to salicylate
-
-
?
S-adenosyl-L-methionine + salicylate
S-adenosyl-L-homocysteine + methyl salicylate
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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Na+, NH4+ and Mg2+ have no effect on isoform SAMT1 activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Mg2+
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5 mM, mild inhibitory effect, less than 35% reduced activity
Na+
5 mM, inhibits SAMT activity less than 30%
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alamethicin
treatment of detached Arabidopsis thaliana leaves efficiently induced methyl salicylate but not methyl benzoate emission
insect herbivory
feeding by the larvae of the diamondback moth on rosette leaves of intact plants resulted in low methyl salicylate and very low methyl benzoate emission
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.004 - 0.087
S-adenosyl-L-methionine
0.00046 - 0.05
salicylate
0.016 - 0.127
salicylic acid
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.013 - 0.028
S-adenosyl-L-methionine
0.056 - 14
salicylate
0.011 - 0.07
salicylic acid
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4
salicylate
Clarkia breweri
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wild type enzyme, pH 7.5, 25C
617
0.4 - 4.3
salicylic acid
765
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000126
salicylic acid, pH 7.5, 20C
0.000228
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crude extract, pH 7.5, 20C
0.00069
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DE-53 DEAE chromatography, pH 7.5, 20C
0.00087
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HAP chromatography, pH 7.5, 20C
0.009
purified recombinant enzyme, pH and temperature not specified in the publication
0.02118
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Mono-Q chromatography, pH 7.5, 20C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
with 8090% of maximum activity
6 - 9
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at pH 6.0 activity is 80% of optimal activity, at pH 8.0 it is 75%, and at pH 9 it is less than 50%
7 - 8
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about 90% activity at pH 7.0, about 40% activity at pH 8.0
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 40
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about 80% activity at 10C, about 90% activity at 20C, about 78% activity at 30C, about 70% activity at 40C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.67
Lillium hybrid cultivar
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
Expression of SAMT mRNA about half the level in petal, where lowest level of mRNA expression is observed
Manually annotated by BRENDA team
expression of SAMT only 2fold higher than in pistil, where lowest level of mRNA expression is observed
Manually annotated by BRENDA team
additional information
little or no AtBSMT1 transcripts observed in roots or siliques of plants
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
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monomer after Mono-Q chromatography
40300
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calculated from sequence
41000
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SDS-PAGE
43360
AlBSMT1 subunit, calculated from sequence
43370
AtBSMT1 subunit, calculated from sequence
74500
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native protein
107000
with a calibrated Sephacryl 200-HR column
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
Lillium hybrid cultivar
x * 41050, about, sequence calculation
dimer
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2 * 40300, calculated from sequence
homodimer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
overall structure of SAMT monomer consists of a globular domain containing the extended beta-sheet characteristic of other SAM-dependent methyltransferases and a unique alpha-helical cap that forms the top one-third of the active site cavity; SAMT crystallized from ammonium sulfate solution, model spanned the entire 359 residues of full-length Clarkia SAMT
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
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SAMT is 100% stable for 30 min at 20C and 80% stable for 30 min at 30C, but after 30 min incubation at 40C it completely loses activity
42
enzyme is stable at temperatures up to 42C for 30 min but 30-min incubation at 65C leads to a 70% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme is active in both Tris- and phosphatecitrate-based buffers, although its activity in phosphate-citrate buffer is about 20% lower than in Tris-buffer
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
the recombinant protein is catalytically stable for more than a year when stored at -80C
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by DE53 DEAE anion exchange, HAP and Mono-Q anion exchange chromatography
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by nickel-based affinity chromatography
N-terminal polyhistidine-tagged SAMT protein purified by Ni2+ affinity chromatography and gel filtration chromatography using a Superdex-75 column
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recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) Codon Plus by metal affinity chromatography
to near homogeneity in two steps, first using anion-exchange column diethylaminoethyl cellulose followed by Mono-Q anion-exchange chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Clarkia breweri SAMT cloned into expression vector pET28a(+) and construct transformed into Escherichia coli BL21(DE3) cells
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coding regions of AlBSMT1 ligated into pCRT7/CT-TOPO TA vector for functional expression in Escherichia coli
coding regions of AtBSMT1 ligated into pCRT7/CT-TOPO TA vector for functional expression in Escherichia coli
expressed in Escherichia coli BL21(DE3) CodonPlus cells
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expressed in Escherichia coli BL21-AI cells
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gene LiBSMT, DNA and amino acid sequence determination and analysis, sequence comparison and phylogenetic analysis and tree, quantitative real-time PCR expression analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Lillium hybrid cultivar
gene NbSAMT, quantitative real-time PCR enzyme expression analysis
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gene PbBSMT, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) codon plus
SAMT cDNA expressed with pET-11a/pET-28 vector in Escherichia coli which synthesizes a functional non-tagged/His-tagged SAMT protein
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subcloned into expression vector pET-28a, containing an N-terminal polyhistidine 6-His-tag, and expressed in Escherichia coli
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
isoform SAMT1 expression drops off to essentially undetectable levels by the time fruits are fully ripened
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leaf wounding and uprooting causes AtBSMT1 transcripts to accumulate quickly within 2 h after treatment with uprooting being more effective; methyl jasmonate treatment, in which plants are exposed to methyl jasmonate vapors in a closed container, results in slow induction
methyljasmonate enhances tomato defense responses prior to pathogen challenge by including production phenolic compounds following gene expression of enzyme involved in phenolic compound production, including benzoic/salicylic acid carboxyl methyltransferase, 1.6fold upregulation of the salicylic acid methylation gene BSMT
physical wounding and 1 mM methyl jasmonate as well as biotic and abiotic stresses (infection with Pseudomonas syringae pv. tabaci or treatment with 1 mM salicylic acid and 1,2-benzisothiazole-1,1-dioxide) induce isoform SAMT1 expression
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under normal growing conditions AtBSMT1 promoter is only active in leaves at the base of the trichomes, in the hydathodes, and in the upper part of the sepals of flowers. Strong AtBSMT1 promoter activity is observed around lesions in the leaves caused by thrips of the genus Frankliniella
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Y147S
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reduced methylation of benzoic acid and 3-hydroxybenzoic acid but slightly increased activity towards jasmonic acid
Y147S/M150H
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significant increase in the ability to turn over jasmonic acid and vanillic acid
Y147S/M150H/F347Y
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slightly increased activity towards short-chain carboxylic acids and jasmonic acid
Y147S/M150H/F347Y/N349I
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highly increased activity towards short-chain carboxylic acids and aromatic acids like hydroxybenzoic acids, vanillic acid, jasmonic acid, cinnamic acid, 4-coumaric acid and caffeic acid, broadest substrate spectrum
Y147S/M150H/I225Q/F347Y
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greatest specific activities against 3-hydroxybenzoic acid, vanillic acid and jasmonic acid but reduced activity towards salicylic acid
additional information
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silencing of gene NbSAMT, quantitative real-time PCR defense-related genes expression analysis
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
priming of tomato seeds with 0.1 mM methyljasmonate before sowing enables the seedlings grown from these seeds to reduce the attack of the soil-borne fungal pathogen Fusarium oxysporum f.sp. lycopersici