Protein Variants | Comment | Organism |
---|---|---|
H398Q | low activity for catalysis of reductive deacylation of (S)-3-hydroxy-3-methylglutaryl-CoA or oxidative acylation of mevaldehyde, but readily catalyzed mevaldehyde reduction | Haloferax volcanii |
additional information | mutant forms of the 403 residue Haloferax volcanii enzyme are constructed to model phosphorylation and infer whether attenuated activity involves interaction with His398. Chimeric Haloferax volcanii-hamster enzymes constructed in an effort to create an active, phosphorylatable chimeric enzyme are inactive or not phosphorylated. Asp is added at position 404 to mimic the introduction of negative charge that would accompany phosphorylation. Enzyme 404D/H398Q was inactive for reductive deacylation of (S)-3-hydroxy-3-methylglutaryl-CoA or oxidative acylation of mevaldehyde, but catalyzed reaction mevaldehyde reduction at 35% the wild-type rate. These observations are consistent with the model that attenuation of catalytic activity results from an ionic interaction between the imidazolium cation of His 398 and the carboxylate anion of Asp404 | Haloferax volcanii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
(R)-mevaldehyde | pH 7.3, 37°C, second step of the reaction, mutant enzyme 404N with Asp is added at position 404 | Haloferax volcanii | |
0.032 | - |
(R)-mevaldehyde | pH 7.3, 37°C, second step of the reaction, wild-type enzyme | Haloferax volcanii | |
0.06 | - |
(S)-3-hydroxy-3-methylglutaryl-CoA | pH 7.3, 37°C, first step of the reaction, wild-type enzyme | Haloferax volcanii | |
0.066 | - |
NADPH | pH 7.3, 37°C, cosubstrate: (S)-3-hydroxy-3-methylglutaryl-CoA, first step of the reaction, wild-type enzyme | Haloferax volcanii | |
0.068 | - |
NADPH | pH 7.3, 37°C, cosubstrate: (S)-3-hydroxy-3-methylglutaryl-CoA, first step of the reaction, mutant enzyme 404D with Asp is added at position 404 | Haloferax volcanii | |
0.068 | - |
NADPH | pH 7.3, 37°C, cosubstrate: (S)-3-hydroxy-3-methylglutaryl-CoA, first step of the reaction, mutant enzyme 404D/H398Q with Asp is added at position 404 | Haloferax volcanii | |
0.072 | - |
NADPH | pH 7.3, 37°C, cosubstrate: (S)-3-hydroxy-3-methylglutaryl-CoA, first step of the reaction, mutant enzyme 404N with Asp is added at position 404 | Haloferax volcanii | |
0.077 | - |
(S)-3-hydroxy-3-methylglutaryl-CoA | pH 7.3, 37°C, first step of the reaction, mutant enzyme 404N with Asp is added at position 404 | Haloferax volcanii | |
0.083 | - |
(S)-3-hydroxy-3-methylglutaryl-CoA | pH 7.3, 37°C, first step of the reaction, mutant enzyme 404D with Asp is added at position 404 | Haloferax volcanii | |
0.083 | - |
(S)-3-hydroxy-3-methylglutaryl-CoA | pH 7.3, 37°C, first step of the reaction, mutant enzyme 404D/H398Q with Asp is added at position 404 | Haloferax volcanii | |
0.084 | - |
(R)-mevaldehyde | pH 7.3, 37°C, second step of the reaction, mutant enzyme 404D/H398Q with Asp is added at position 404 | Haloferax volcanii | |
0.1 | - |
(R)-mevaldehyde | pH 7.3, 37°C, second step of the reaction, mutant enzyme H398Q | Haloferax volcanii | |
0.177 | - |
NADPH | pH 7.3, 37°C, cosubstrate: (S)-3-hydroxy-3-methylglutaryl-CoA, first step of the reaction, mutant enzyme H398Q | Haloferax volcanii | |
0.2 | - |
(S)-3-hydroxy-3-methylglutaryl-CoA | pH 7.3, 37°C, first step of the reaction, mutant enzyme H398Q | Haloferax volcanii | |
0.55 | - |
NADPH | pH 7.3, 37°C, cosubstrate: mevaldehyde, second step of the reaction, wild-type enzyme | Haloferax volcanii | |
2.1 | - |
NADPH | pH 7.3, 37°C, cosubstrate: mevaldehyde, second step of the reaction, mutant enzyme 404D/H398Q with Asp is added at position 404 | Haloferax volcanii | |
3 | - |
NADPH | pH 7.3, 37°C, cosubstrate: mevaldehyde, second step of the reaction, mutant enzyme H398Q | Haloferax volcanii | |
4.2 | - |
NADPH | pH 7.3, 37°C, cosubstrate: mevaldehyde, second step of the reaction, mutant enzyme 404N with Asp is added at position 404 | Haloferax volcanii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haloferax volcanii | Q59468 | - |
- |
Haloferax volcanii DSM 3757 | Q59468 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Haloferax volcanii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-mevaldehyde + NADPH + H+ | second step of the reaction | Haloferax volcanii | (R)-mevalonate + NADP+ | - |
? | |
(R)-mevaldehyde + NADPH + H+ | second step of the reaction | Haloferax volcanii DSM 3757 | (R)-mevalonate + NADP+ | - |
? | |
(S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADPH + 2 H+ | overall reaction | Haloferax volcanii | (R)-mevalonate + CoA + 2 NADP+ | - |
? | |
(S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADPH + 2 H+ | overall reaction | Haloferax volcanii DSM 3757 | (R)-mevalonate + CoA + 2 NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxy-3-methylglutaryl-coenzyme A reductase | - |
Haloferax volcanii |
HMG-CoA reductase | - |
Haloferax volcanii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Haloferax volcanii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.3 | - |
assay at | Haloferax volcanii |