Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain NM522 | Dickeya chrysanthemi |
Crystallization (Comment) | Organism |
---|---|
the inactive D178A PME mutant in complex with specifically methylated hexagalacturonates, 3 mg/ml protein, crystallization solutions contain 0.1 M MES, pH 6.5, 10% dioxane and 1.6 M ammonium sulfate, and dilutions of that crystallisation buffer with H2O, or 0.1 M MES, pH 6.5, and 12% w/v PEG 20000, X-ray diffraction structure determination and anaylsis at 1.7-1.9 A resolution | Dickeya chrysanthemi |
Protein Variants | Comment | Organism |
---|---|---|
D178A | site-directed mutagenesis, inactive mutant | Dickeya chrysanthemi |
D199A | site-directed mutagenesis, inactive mutant | Dickeya chrysanthemi |
M306A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Dickeya chrysanthemi |
Q153A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Dickeya chrysanthemi |
Q177A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Dickeya chrysanthemi |
R267A | site-directed mutagenesis, inactive mutant | Dickeya chrysanthemi |
T272A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Dickeya chrysanthemi |
V198A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Dickeya chrysanthemi |
W269A | site-directed mutagenesis, inactive mutant | Dickeya chrysanthemi |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Polygalacturonate | completely deesterified pectin, a competitive inhibitor of PME | Dickeya chrysanthemi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics, kinetic mechanism | Dickeya chrysanthemi | |
0.06 | - |
Pectin | mutant V198A, pH 7.0, 30°C | Dickeya chrysanthemi | |
0.13 | - |
Pectin | wild-type enzyme, pH 7.0, 30°C | Dickeya chrysanthemi | |
0.22 | - |
Pectin | mutant Q177A, pH 7.0, 30°C | Dickeya chrysanthemi | |
0.53 | - |
Pectin | mutant M306A, pH 7.0, 30°C | Dickeya chrysanthemi | |
0.77 | - |
Pectin | mutant T272A, pH 7.0, 30°C | Dickeya chrysanthemi | |
2.2 | - |
Pectin | mutant Q153A, pH 7.0, 30°C | Dickeya chrysanthemi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
pectin + H2O | Dickeya chrysanthemi | the enzyme catalyses the essential first step in bacterial invasion of plant tissues | methanol + pectate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dickeya chrysanthemi | - |
a plant pathogen, strain B374 | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
pectin + n H2O = n methanol + pectate | two conserved aspartates are the nucleophile and general acid-base in the reaction mechanism, respectively, the catalytic site is formed by the conserved resisues Asp178, Asp199, and Arg267, molecular basis of the processive action of the enzyme, overview | Dickeya chrysanthemi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | substrate specificity, overview | Dickeya chrysanthemi | ? | - |
? | |
pectin + H2O | the enzyme catalyses the essential first step in bacterial invasion of plant tissues | Dickeya chrysanthemi | methanol + pectate | - |
? | |
pectin + H2O | the enzyme shows a sequential pattern of demethylation due to the preferential binding of methylated sugar residues upstream of the catalytic site, and demethylated residues downstream, which drives the enzyme along the pectin molecule | Dickeya chrysanthemi | methanol + pectate | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme possesses a parallel beta-helix architecture, three dimensional structure of PME, overview | Dickeya chrysanthemi |
Synonyms | Comment | Organism |
---|---|---|
pectin methylesterase | - |
Dickeya chrysanthemi |
PME | - |
Dickeya chrysanthemi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Dickeya chrysanthemi |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
40 | - |
Pectin | mutant Q177A, pH 7.0, 30°C | Dickeya chrysanthemi | |
59 | - |
Pectin | mutant M306A, pH 7.0, 30°C | Dickeya chrysanthemi | |
68 | - |
Pectin | mutant V198A, pH 7.0, 30°C | Dickeya chrysanthemi | |
232 | - |
Pectin | mutant Q153A, pH 7.0, 30°C | Dickeya chrysanthemi | |
425 | - |
Pectin | mutant T272A, pH 7.0, 30°C | Dickeya chrysanthemi | |
450 | - |
Pectin | wild-type enzyme, pH 7.0, 30°C | Dickeya chrysanthemi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Dickeya chrysanthemi |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
16 | - |
Polygalacturonate | - |
Dickeya chrysanthemi |