BRENDA - Enzyme Database
show all sequences of 2.1.1.274

Structural basis for substrate recognition in the salicylic acid carboxyl methyltransferase family

Zubieta, C.; Ross, J.R.; Koscheski, P.; Yang, Y.; Pichersky, E.; Noel, J.P.; Plant Cell 15, 1704-1716 (2003)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
Clarkia breweri SAMT cloned into expression vector pET28a(+) and construct transformed into Escherichia coli BL21(DE3) cells
Clarkia breweri
Crystallization (Commentary)
Crystallization (Commentary)
Organism
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
Clarkia breweri
Engineering
Protein Variants
Commentary
Organism
Y147S
reduced methylation of benzoic acid and 3-hydroxybenzoic acid but slightly increased activity towards jasmonic acid
Clarkia breweri
Y147S/M150H
significant increase in the ability to turn over jasmonic acid and vanillic acid
Clarkia breweri
Y147S/M150H/F347Y
slightly increased activity towards short-chain carboxylic acids and jasmonic acid
Clarkia breweri
Y147S/M150H/F347Y/N349I
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
Clarkia breweri
Y147S/M150H/I225Q/F347Y
greatest specific activities against 3-hydroxybenzoic acid, vanillic acid and jasmonic acid but reduced activity towards salicylic acid
Clarkia breweri
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.023
-
salicylate
wild type enzyme, pH 7.5, 25°C
Clarkia breweri
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41000
-
SDS-PAGE
Clarkia breweri
Organism
Organism
UniProt
Commentary
Textmining
Clarkia breweri
Q9SPV4
-
-
Purification (Commentary)
Purification (Commentary)
Organism
N-terminal polyhistidine-tagged SAMT protein purified by Ni2+ affinity chromatography and gel filtration chromatography using a Superdex-75 column
Clarkia breweri
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
additional information
no measurable methylation of jasmonic acid by wild-type SAMT using concentrations up to 5 mM
723402
Clarkia breweri
?
-
-
-
?
additional information
wild-type is also able to methylate benzoic acid with 48% activity compared to salicylate methylation
723402
Clarkia breweri
?
-
-
-
?
S-adenosyl-L-methionine + 3-hydroxybenzoic acid
17% activity of wild-type enzyme compared to salicylate methylation
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate
-
-
-
?
S-adenosyl-L-methionine + jasmonic acid
Y147S/M150H double mutant and Y147S/M150H/F347Y triple mutant are able to turn over jasmonic acid, while preserving substantial salicylate methylating activity
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl jasmonate
-
-
-
?
S-adenosyl-L-methionine + salicylate
-
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl salicylate
-
-
-
?
S-adenosyl-L-methionine + vanillic acid
5.1% activity of wild-type enzyme compared to salicylate methylation
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl 4-hydroxy-3-methoxybenzoate
-
-
-
?
Subunits
Subunits
Commentary
Organism
homodimer
2 * 41000
Clarkia breweri
Synonyms
Synonyms
Commentary
Organism
SA carboxyl methyltransferase
-
Clarkia breweri
SAMT
-
Clarkia breweri
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.092
-
salicylate
wild type enzyme, pH 7.5, 25°C
Clarkia breweri
Cloned(Commentary) (protein specific)
Commentary
Organism
Clarkia breweri SAMT cloned into expression vector pET28a(+) and construct transformed into Escherichia coli BL21(DE3) cells
Clarkia breweri
Crystallization (Commentary) (protein specific)
Crystallization
Organism
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
Clarkia breweri
Engineering (protein specific)
Protein Variants
Commentary
Organism
Y147S
reduced methylation of benzoic acid and 3-hydroxybenzoic acid but slightly increased activity towards jasmonic acid
Clarkia breweri
Y147S/M150H
significant increase in the ability to turn over jasmonic acid and vanillic acid
Clarkia breweri
Y147S/M150H/F347Y
slightly increased activity towards short-chain carboxylic acids and jasmonic acid
Clarkia breweri
Y147S/M150H/F347Y/N349I
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
Clarkia breweri
Y147S/M150H/I225Q/F347Y
greatest specific activities against 3-hydroxybenzoic acid, vanillic acid and jasmonic acid but reduced activity towards salicylic acid
Clarkia breweri
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.023
-
salicylate
wild type enzyme, pH 7.5, 25°C
Clarkia breweri
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41000
-
SDS-PAGE
Clarkia breweri
Purification (Commentary) (protein specific)
Commentary
Organism
N-terminal polyhistidine-tagged SAMT protein purified by Ni2+ affinity chromatography and gel filtration chromatography using a Superdex-75 column
Clarkia breweri
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
additional information
no measurable methylation of jasmonic acid by wild-type SAMT using concentrations up to 5 mM
723402
Clarkia breweri
?
-
-
-
?
additional information
wild-type is also able to methylate benzoic acid with 48% activity compared to salicylate methylation
723402
Clarkia breweri
?
-
-
-
?
S-adenosyl-L-methionine + 3-hydroxybenzoic acid
17% activity of wild-type enzyme compared to salicylate methylation
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate
-
-
-
?
S-adenosyl-L-methionine + jasmonic acid
Y147S/M150H double mutant and Y147S/M150H/F347Y triple mutant are able to turn over jasmonic acid, while preserving substantial salicylate methylating activity
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl jasmonate
-
-
-
?
S-adenosyl-L-methionine + salicylate
-
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl salicylate
-
-
-
?
S-adenosyl-L-methionine + vanillic acid
5.1% activity of wild-type enzyme compared to salicylate methylation
723402
Clarkia breweri
S-adenosyl-L-homocysteine + methyl 4-hydroxy-3-methoxybenzoate
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 41000
Clarkia breweri
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.092
-
salicylate
wild type enzyme, pH 7.5, 25°C
Clarkia breweri
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4
-
salicylate
wild type enzyme, pH 7.5, 25°C
Clarkia breweri
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4
-
salicylate
wild type enzyme, pH 7.5, 25°C
Clarkia breweri
Other publictions for EC 2.1.1.274
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
736189
Wang
Cloning and characterization o ...
Lilium hybrid cultivar
Genet. Mol. Res.
14
14510-14521
2015
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2
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736696
Krol
Priming of seeds with methyl j ...
Solanum lycopersicum
J. Plant Physiol.
179
122-132
2015
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1
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736842
Ludwig-Mueller
A novel methyltransferase from ...
Plasmodiophora brassicae
Mol. Plant Pathol.
16
349-364
2015
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736841
Zhu
Salicylic acid and jasmonic ac ...
Nicotiana benthamiana
Mol. Plant Microbe Interact.
27
567-577
2014
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726129
Lin
Overexpression of a soybean sa ...
Glycine max
Plant Biotechnol. J.
11
1135-1145
2013
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1
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1
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725781
Yao
QM/MM free energy simulations ...
Clarkia breweri
J. Phys. Chem. B
115
389-396
2011
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726013
Liu
Altering expression of benzoic ...
Arabidopsis thaliana
Mol. Plant Microbe Interact.
23
82-90
2010
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726172
Tieman
Functional analysis of a tomat ...
Solanum lycopersicum
Plant J.
62
113-123
2010
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726132
Kwon
-
Biotic and abiotic stresses in ...
Atropa belladonna
Plant Biotechnol.
26
207-215
2009
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723402
Zubieta
Structural basis for substrate ...
Clarkia breweri
Plant Cell
15
1704-1716
2003
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723416
Chen
An Arabidopsis thaliana gene f ...
Arabidopsis lyrata, Arabidopsis thaliana
Plant J.
36
577-588
2003
2
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721415
Negre
Novel S-adenosyl-L-methionine: ...
Antirrhinum majus
Arch. Biochem. Biophys.
406
261-270
2002
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721412
Ross
S-Adenosyl-L-methionine:salicy ...
Clarkia breweri
Arch. Biochem. Biophys.
367
9-16
1999
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