Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Vinokur, J.M.; Korman, T.P.; Sawaya, M.R.; Collazo, M.; Cascio, D.; Bowie, J.U.
    Structural analysis of mevalonate-3-kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases (2015), Protein Sci., 24, 212-220.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.185 expression in Escherichia coli Thermoplasma acidophilum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.185 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

EC Number Protein Variants Comment Organism
2.7.1.185 L18A kcat/KM for (R)-mevalonate is 4.4% compared to the wild-type value Thermoplasma acidophilum
2.7.1.185 R185A mutation results in no detectable activity Thermoplasma acidophilum
2.7.1.185 R185K kcat/KM for (R)-mevalonate is 0.5% compared to the wild-type value Thermoplasma acidophilum
2.7.1.185 S105A kcat/KM for (R)-mevalonate is 10.3% compared to the wild-type value Thermoplasma acidophilum
2.7.1.185 T275A kcat/KM for (R)-mevalonate is 25.6% compared to the wild-type value Thermoplasma acidophilum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.185 0.13
-
 (R)-mevalonate pH 8.5, 55°C, wild-type enzyme Thermoplasma acidophilum
2.7.1.185 0.27
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme S105A Thermoplasma acidophilum
2.7.1.185 0.59
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme T275A Thermoplasma acidophilum
2.7.1.185 1.68
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme R185K Thermoplasma acidophilum
2.7.1.185 3.1
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme L18A Thermoplasma acidophilum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.185 ATP + (R)-mevalonate Thermoplasma acidophilum
-
 ADP + (R)-3-phosphomevalonate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.185 Thermoplasma acidophilum Q9HIN1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.185
-
 Thermoplasma acidophilum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.185 ATP + (R)-mevalonate
-
 Thermoplasma acidophilum ADP + (R)-3-phosphomevalonate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.185 dimer the monomeric and dimeric forms have equal activity under optimal conditions Thermoplasma acidophilum
2.7.1.185 monomer the monomeric and dimeric forms have equal activity under optimal conditions Thermoplasma acidophilum

Synonyms

EC Number Synonyms Comment Organism
2.7.1.185 M3K
-
 Thermoplasma acidophilum
2.7.1.185 Ta1305 locus name Thermoplasma acidophilum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.185 55
-
 the monomeric and dimeric forms have equal activity under optimal conditions Thermoplasma acidophilum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.185 0.31
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme R185K Thermoplasma acidophilum
2.7.1.185 1.16
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme S105A Thermoplasma acidophilum
2.7.1.185 5.1
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme L18A Thermoplasma acidophilum
2.7.1.185 5.3
-
 (R)-mevalonate pH 8.5, 55°C, wild-type enzyme Thermoplasma acidophilum
2.7.1.185 5.8
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme T275A Thermoplasma acidophilum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.185 8.5
-
 the monomeric and dimeric forms have equal activity under optimal conditions Thermoplasma acidophilum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.1.185 0.19
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme R185K Thermoplasma acidophilum
2.7.1.185 1.7
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme L18A Thermoplasma acidophilum
2.7.1.185 4
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme S105A Thermoplasma acidophilum
2.7.1.185 10
-
 (R)-mevalonate pH 8.5, 55°C, mutant enzyme T275A Thermoplasma acidophilum
2.7.1.185 39
-
 (R)-mevalonate pH 8.5, 55°C, wild-type enzyme Thermoplasma acidophilum