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Literature summary for 2.1.1.274 extracted from

  • Zubieta, C.; Ross, J.R.; Koscheski, P.; Yang, Y.; Pichersky, E.; Noel, J.P.
    Structural basis for substrate recognition in the salicylic acid carboxyl methyltransferase family (2003), Plant Cell, 15, 1704-1716.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.1.1.274

Cloned(Commentary)

Cloned (Comment) Organism
Clarkia breweri SAMT cloned into expression vector pET28a(+) and construct transformed into Escherichia coli BL21(DE3) cells Clarkia breweri

Crystallization (Commentary)

Crystallization (Comment) 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 Clarkia breweri
SAMT crystallized from ammonium sulfate solution, model spanned the entire 359 residues of full-length Clarkia SAMT Clarkia breweri

Protein Variants

Protein Variants Comment 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 Comment 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] Comment Organism
41000
-
 SDS-PAGE Clarkia breweri

Organism

Organism UniProt Comment Textmining
Clarkia breweri Q9SPV4
-
-

Purification (Commentary)

Purification (Comment) 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information no measurable methylation of jasmonic acid by wild-type SAMT using concentrations up to 5 mM Clarkia breweri ?
-
 ?
additional information wild-type is also able to methylate benzoic acid with 48% activity compared to salicylate methylation Clarkia breweri ?
-
 ?
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
-
 ?
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
-
 ?
S-adenosyl-L-methionine + salicylate
-
 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 Clarkia breweri S-adenosyl-L-homocysteine + methyl 4-hydroxy-3-methoxybenzoate
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 41000 Clarkia breweri

Synonyms

Synonyms Comment Organism
SA carboxyl methyltransferase
-
 Clarkia breweri
SAMT
-
 Clarkia breweri

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment 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 Comment Organism Structure
4
-
 salicylate wild type enzyme, pH 7.5, 25°C Clarkia breweri