EC Number | Cloned (Comment) | Organism |
---|---|---|
2.1.1.274 | Clarkia breweri SAMT cloned into expression vector pET28a(+) and construct transformed into Escherichia coli BL21(DE3) cells | Clarkia breweri |
2.1.1.278 | expression in Escherichia coli | Arabidopsis thaliana |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
2.1.1.274 | 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 | Clarkia breweri |
2.1.1.274 | SAMT crystallized from ammonium sulfate solution, model spanned the entire 359 residues of full-length Clarkia SAMT | Clarkia breweri |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.1.1.274 | Y147S | reduced methylation of benzoic acid and 3-hydroxybenzoic acid but slightly increased activity towards jasmonic acid | Clarkia breweri |
2.1.1.274 | Y147S/M150H | significant increase in the ability to turn over jasmonic acid and vanillic acid | Clarkia breweri |
2.1.1.274 | Y147S/M150H/F347Y | slightly increased activity towards short-chain carboxylic acids and jasmonic acid | Clarkia breweri |
2.1.1.274 | 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 |
2.1.1.274 | Y147S/M150H/I225Q/F347Y | greatest specific activities against 3-hydroxybenzoic acid, vanillic acid and jasmonic acid but reduced activity towards salicylic acid | Clarkia breweri |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.1.274 | 0.023 | - |
salicylate | wild type enzyme, pH 7.5, 25°C | Clarkia breweri | |
2.1.1.278 | 0.013 | - |
(indol-3-yl)acetate | pH 7.5, 25°C | Arabidopsis thaliana |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
2.1.1.278 | Mg2+ | binds Mg2+ | Arabidopsis thaliana |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.1.1.274 | 41000 | - |
SDS-PAGE | Clarkia breweri |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.1.1.278 | S-adenosyl-L-methionine + (indol-3-yl)acetate | Arabidopsis thaliana | - |
S-adenosyl-L-homocysteine + methyl (indol-3-yl)acetate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.1.1.274 | Clarkia breweri | Q9SPV4 | - |
- |
2.1.1.278 | Arabidopsis thaliana | Q9FLN8 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.1.1.274 | N-terminal polyhistidine-tagged SAMT protein purified by Ni2+ affinity chromatography and gel filtration chromatography using a Superdex-75 column | Clarkia breweri |
2.1.1.278 | - |
Arabidopsis thaliana |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.1.1.274 | additional information | no measurable methylation of jasmonic acid by wild-type SAMT using concentrations up to 5 mM | Clarkia breweri | ? | - |
? | |
2.1.1.274 | additional information | wild-type is also able to methylate benzoic acid with 48% activity compared to salicylate methylation | Clarkia breweri | ? | - |
? | |
2.1.1.274 | S-adenosyl-L-methionine + 3-hydroxybenzoic acid | 17% activity of wild-type enzyme compared to salicylate methylation | Clarkia breweri | S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate | - |
? | |
2.1.1.274 | 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 | Clarkia breweri | S-adenosyl-L-homocysteine + methyl jasmonate | - |
? | |
2.1.1.274 | S-adenosyl-L-methionine + salicylate | - |
Clarkia breweri | S-adenosyl-L-homocysteine + methyl salicylate | - |
? | |
2.1.1.274 | S-adenosyl-L-methionine + vanillic acid | 5.1% activity of wild-type enzyme compared to salicylate methylation | Clarkia breweri | S-adenosyl-L-homocysteine + methyl 4-hydroxy-3-methoxybenzoate | - |
? | |
2.1.1.278 | S-adenosyl-L-methionine + (indol-3-yl)acetate | - |
Arabidopsis thaliana | S-adenosyl-L-homocysteine + methyl (indol-3-yl)acetate | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.1.1.274 | homodimer | 2 * 41000 | Clarkia breweri |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.1.1.274 | SA carboxyl methyltransferase | - |
Clarkia breweri |
2.1.1.274 | SAMT | - |
Clarkia breweri |
2.1.1.278 | At5g55250 | - |
Arabidopsis thaliana |
2.1.1.278 | IAA carboxylmethyltransferase | - |
Arabidopsis thaliana |
2.1.1.278 | IAMT | - |
Arabidopsis thaliana |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.1.274 | 0.092 | - |
salicylate | wild type enzyme, pH 7.5, 25°C | Clarkia breweri | |
2.1.1.278 | 0.028 | - |
(indol-3-yl)acetate | pH 7.5, 25°C | Arabidopsis thaliana |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.1.274 | 4 | - |
salicylate | wild type enzyme, pH 7.5, 25°C | Clarkia breweri | |
2.1.1.278 | 2.15 | - |
(indol-3-yl)acetate | pH 7.5, 25°C | Arabidopsis thaliana |