Cloned (Comment) | Organism |
---|---|
gene SABATH4, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, overview, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Populus trichocarpa |
Protein Variants | Comment | Organism |
---|---|---|
M156H | site-directed mutagenesis, the mutation of Met156 to His results in a switch from a preference for salicylic acid (SA) over benzoic acid (BA) in wild-type PtSABATH4 to a preference for BA over SA in the M156H mutant | Populus trichocarpa |
M314V | site-directed mutagenesis, the mutation results in decreased enzymatic activities towards both the substrates salicylic acid (SA) over benzoic acid (BA), but not in a substrate switch | Populus trichocarpa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + benzoate | Populus trichocarpa | - |
methyl benzoate + S-adenosyl-L-homocysteine | - |
? | |
S-adenosyl-L-methionine + salicylate | Populus trichocarpa | - |
methyl salicylate + S-adenosyl-L-homocysteine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Populus trichocarpa | A9PF83 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Populus trichocarpa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.95 | - |
below, recombinant wild-type PtSABATH4 with benzoic acid, pH 7.4, 25°C | Populus trichocarpa |
27.96 | - |
recombinant wild-type PtSABATH4 with salicylic acid, pH 7.4, 25°C | Populus trichocarpa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | enzyme PtSABATH4 exhibits high enzymatic activity towards the substrate salicylate (SA) and weak activity towards benzoic acid, jasmonic acid, and farnesoic acid. PtSABATH4 does not show any activity towards indole-3-acetic acid, vanillic acid, nicotinic acid, coumalic acid, and trans-cinnamic acid. PtSABATH4 shows at least 4.3fold higher enzymatic activity towards the substrate SA. PtSABATH4 displays the highest level of catalytic activity towards SA and a relatively low level of activity towards BA. Preference for salicylic acid (SA) over benzoic acid (BA) in wild-type PtSABATH4 and preference for BA over SA in the PtSABATH4 M156H mutant | Populus trichocarpa | ? | - |
- |
|
S-adenosyl-L-methionine + benzoate | - |
Populus trichocarpa | methyl benzoate + S-adenosyl-L-homocysteine | - |
? | |
S-adenosyl-L-methionine + salicylate | - |
Populus trichocarpa | methyl salicylate + S-adenosyl-L-homocysteine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
class II SABATH protein | - |
Populus trichocarpa |
More | see also EC 2.1.1.273 | Populus trichocarpa |
PtSABATH4 | - |
Populus trichocarpa |
S-adenosyl-L-methionine:salicylic acid carboxyl methyltransferase | UniProt | Populus trichocarpa |
SABATH methyltransferase 4 | UniProt | Populus trichocarpa |
SABATH4 | - |
Populus trichocarpa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Populus trichocarpa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Populus trichocarpa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Populus trichocarpa |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the SABATH family, phylogenetic analysis and tree, detailed overview. Twenty-eight Populus SABATH genes are divided into three classes with distinct divergences in their gene structure, expression responses to abiotic stressors and enzymatic properties of encoded proteins. Populus class I SABATH proteins convert indole-3-acetic acid (IAA) to methyl-IAA, class II SABATH proteins convert benzoic acid (BA) and salicylic acid (SA) to methyl-BA and methyl-SA, while class III SABATH proteins convert farnesoic acid (FA) to methyl-FA. For Populus class II SABATH proteins, both forward and reverse mutagenesis studies show that a single amino acid switch between PtSABATH4 and PtSABATH24 results in substrate switch. Of the Populus SABATH class II proteins, PtSABATH4 and 24 show the highest activity towards SA and BA, respectively | Populus trichocarpa |
malfunction | for Populus class II SABATH proteins, both forward and reverse mutagenesis studies show that a single amino acid switch between PtSABATH4 and PtSABATH24 results in substrate switch. The mutation of Met156 to His results in a switch from a preference for salicylic acid (SA) over benzoic acid (BA) in wild-type PtSABATH4 to a preference for BA over SA in the M156H mutant. The mutation of His157 of PtSABATH24 (EC 2.1.1.273) to a methionine residue also results in a switch from a preference for BA over SA in wild-type PtSABATH24 to a preference for SA over BA in the H157M mutant. The PtSABATH4 mutation M314V results in decreased enzymatic activities towards both the substrates salicylic acid (SA) over benzoic acid (BA), but not in a substrate switch | Populus trichocarpa |
metabolism | expression patterns of Populus SABATH genes under normal growth conditions and abiotic stress, overview | Populus trichocarpa |
additional information | three-dimensional structure modeling of PtSABATH4 based on the 1M6E crystal structure. Similar to 1M6E, residues Met156 and Met314 of PtSABATH4 also create a molecular clamp that encompasses the benzyl ring of salicylate | Populus trichocarpa |