Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Thermoplasma acidophilum |
Crystallization (Comment) | Organism |
---|---|
hanging drop method, crystal structure of mevalonate-3-kinase in the apo form, and with bound substrates is determined and compared to mevalonate diphosphate decarboxylase structures. The crystal structure of mevalonate-3-kinase provides insight into the mechanism of mevalonate diphosphate decarboxylase. Despite sharing nearly identical overall folds, important active site differences are identified. Glu140 in the center of the mevalonate-3-kinase active site is responsible for binding mevalonate while excluding mevalonate 5-diphosphate, Arg185/Ser105 catalyze phosphate transfer, and an invariant Asp/Lys pair previously thought to be responsible for phosphorylation in mevalonate diphosphate decarboxylase, is missing in mevalonate-3-kinase and replaced by non-essential Thr275/Leu18. A model is proposed in which mevalonate-3-kinase and mevalonate diphosphate decarboxylase both phosphorylate by stabilizing a phosphotransfer transition state (mevalonate-3-kinase via Arg185/Ser105, mevalonate diphosphate decarboxylase via Lys188), suggesting the invariant Asp/Lys pair unique to mevalonate diphosphate decarboxylase may be critical for the decarboxylation step rather than phosphorylation | Thermoplasma acidophilum |
Protein Variants | Comment | Organism |
---|---|---|
L18A | kcat/KM for (R)-mevalonate is 4.4% compared to the wild-type value | Thermoplasma acidophilum |
R185A | mutation results in no detectable activity | Thermoplasma acidophilum |
R185K | kcat/KM for (R)-mevalonate is 0.5% compared to the wild-type value | Thermoplasma acidophilum |
S105A | kcat/KM for (R)-mevalonate is 10.3% compared to the wild-type value | Thermoplasma acidophilum |
T275A | kcat/KM for (R)-mevalonate is 25.6% compared to the wild-type value | Thermoplasma acidophilum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.13 | - |
(R)-mevalonate | pH 8.5, 55°C, wild-type enzyme | Thermoplasma acidophilum | |
0.27 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme S105A | Thermoplasma acidophilum | |
0.59 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme T275A | Thermoplasma acidophilum | |
1.68 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme R185K | Thermoplasma acidophilum | |
3.1 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme L18A | Thermoplasma acidophilum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (R)-mevalonate | Thermoplasma acidophilum | - |
ADP + (R)-3-phosphomevalonate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermoplasma acidophilum | Q9HIN1 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Thermoplasma acidophilum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (R)-mevalonate | - |
Thermoplasma acidophilum | ADP + (R)-3-phosphomevalonate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | the monomeric and dimeric forms have equal activity under optimal conditions | Thermoplasma acidophilum |
monomer | the monomeric and dimeric forms have equal activity under optimal conditions | Thermoplasma acidophilum |
Synonyms | Comment | Organism |
---|---|---|
M3K | - |
Thermoplasma acidophilum |
Ta1305 | locus name | Thermoplasma acidophilum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
the monomeric and dimeric forms have equal activity under optimal conditions | Thermoplasma acidophilum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.31 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme R185K | Thermoplasma acidophilum | |
1.16 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme S105A | Thermoplasma acidophilum | |
5.1 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme L18A | Thermoplasma acidophilum | |
5.3 | - |
(R)-mevalonate | pH 8.5, 55°C, wild-type enzyme | Thermoplasma acidophilum | |
5.8 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme T275A | Thermoplasma acidophilum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
the monomeric and dimeric forms have equal activity under optimal conditions | Thermoplasma acidophilum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.19 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme R185K | Thermoplasma acidophilum | |
1.7 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme L18A | Thermoplasma acidophilum | |
4 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme S105A | Thermoplasma acidophilum | |
10 | - |
(R)-mevalonate | pH 8.5, 55°C, mutant enzyme T275A | Thermoplasma acidophilum | |
39 | - |
(R)-mevalonate | pH 8.5, 55°C, wild-type enzyme | Thermoplasma acidophilum |