Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | selection of functional mALAS2 variants from the Thr148/Asn150 library of constructs is accomplished by reversing the 5-aminolevulinate auxotrophic phenotype of Escherichia coli hemA (HU227) cells | Mus musculus |
N150A | site-directed mutagenesis, the mutation significantly reduces the rate of quinonoid intermediate formation in the forward direction | Mus musculus |
N150F | site-directed mutagenesis, the mutation significantly reduces the rate of quinonoid intermediate formation in the forward direction, while increasing the reverse reaction rate | Mus musculus |
N150G | site-directed mutagenesis, the mutation significantly reduces the rate of quinonoid intermediate formation in the forward direction | Mus musculus |
N150H | site-directed mutagenesis, the mutation significantly reduces the rate of quinonoid intermediate formation in the forward direction, while increasing the reverse reaction rate | Mus musculus |
N150W | site-directed mutagenesis, the mutation significantly reduces the rate of quinonoid intermediate formation in the forward direction | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | pre-steady-state and steady-state kinetics of wild-type and mutant enzymes, stopped-flow measurements, single and mutiple turnover rates, equilibrium dissociation constants, binding isotherms, detailed overview | Mus musculus | |
0.0011 | - |
succinyl-CoA | mutant N150W, pH 7.5, 18°C | Mus musculus | |
0.0012 | - |
succinyl-CoA | mutant N150H, pH 7.5, 18°C | Mus musculus | |
0.0021 | - |
succinyl-CoA | mutant N150G, pH 7.5, 18°C | Mus musculus | |
0.0022 | - |
succinyl-CoA | mutant N150F, pH 7.5, 18°C | Mus musculus | |
0.0023 | - |
succinyl-CoA | wild-type enzyme, pH 7.5, 18°C | Mus musculus | |
0.0038 | - |
succinyl-CoA | mutant N150A, pH 7.5, 18°C | Mus musculus | |
2 | 3 | glycine | wild-type enzyme, pH 7.5, 18°C | Mus musculus | |
11 | - |
glycine | mutant N150G, pH 7.5, 18°C | Mus musculus | |
12 | - |
glycine | mutant N150F, pH 7.5, 18°C | Mus musculus | |
16 | - |
glycine | mutant N150A, pH 7.5, 18°C | Mus musculus | |
16 | - |
glycine | mutant N150H, pH 7.5, 18°C | Mus musculus | |
39 | - |
glycine | mutant N150W, pH 7.5, 18°C | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
succinyl-CoA + glycine | Mus musculus | - |
5-aminolevulinate + CoA + CO2 | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | P08680 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes | Mus musculus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
succinyl-CoA + glycine = 5-aminolevulinate + CoA + CO2 | proposed chemical mechanism of enzyme ALAS2 via (I) internal aldimine complex, (II) glycine-external aldimine, (III) quinonoid intermediate I, (IV) glycine-succinyl-CoA condensation intermediate, (V) 2-amino-3-ketoadipate intermediate, (VI) enol intermediate, (VII) quinonoid intermediate II, and (VIII) 5-aminolevulinate-external aldimine | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the wild-type enzyme catalyzes the conversion of 5-aminolevulinate into the quinonoid intermediate at a rate 6.3fold slower than the formation of the same quinonoid intermediate from glycine and succinyl-CoA. The mutant N150F enzyme catalyzes the forward reaction at a 1.2fold faster rate than that of the reverse reaction, and the N150H variant reverses the rate values with a 1.7fold faster rate for the reverse reaction than that for the forward reaction. Steric constraints imposed by the active site of wild-type enzyme ALAS contribute toward the amino acid substrate specificity | Mus musculus | ? | - |
? | |
succinyl-CoA + glycine | - |
Mus musculus | 5-aminolevulinate + CoA + CO2 | - |
r |
Synonyms | Comment | Organism |
---|---|---|
ALAS2 | - |
Mus musculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
18 | 37 | assay at | Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
succinyl-CoA | mutant N150H, pH 7.5, 18°C | Mus musculus | |
0.06 | - |
succinyl-CoA | mutant N150F, pH 7.5, 18°C | Mus musculus | |
0.09 | - |
succinyl-CoA | mutant N150W, pH 7.5, 18°C | Mus musculus | |
0.13 | - |
succinyl-CoA | mutant N150A, pH 7.5, 18°C | Mus musculus | |
0.15 | - |
succinyl-CoA | mutant N150G, pH 7.5, 18°C | Mus musculus | |
0.16 | - |
succinyl-CoA | wild-type enzyme, pH 7.5, 18°C | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on | Mus musculus |
General Information | Comment | Organism |
---|---|---|
metabolism | 5-aminolevulinate synthase catalyzes the initial step of mammalian heme biosynthesis | Mus musculus |
additional information | residue Asn150 is essential for establishing a catalytic balance between the forward and reverse reactions | Mus musculus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0023 | - |
glycine | mutant N150W, pH 7.5, 18°C | Mus musculus | |
0.0025 | - |
glycine | mutant N150H, pH 7.5, 18°C | Mus musculus | |
0.005 | - |
glycine | mutant N150F, pH 7.5, 18°C | Mus musculus | |
0.007 | - |
glycine | wild-type enzyme, pH 7.5, 18°C | Mus musculus | |
0.0081 | - |
glycine | mutant N150A, pH 7.5, 18°C | Mus musculus | |
0.0136 | - |
glycine | mutant N150G, pH 7.5, 18°C | Mus musculus | |
27 | - |
succinyl-CoA | mutant N150F, pH 7.5, 18°C | Mus musculus | |
33 | - |
succinyl-CoA | mutant N150H, pH 7.5, 18°C | Mus musculus | |
34 | - |
succinyl-CoA | mutant N150A, pH 7.5, 18°C | Mus musculus | |
70 | - |
succinyl-CoA | wild-type enzyme, pH 7.5, 18°C | Mus musculus | |
71 | - |
succinyl-CoA | mutant N150G, pH 7.5, 18°C | Mus musculus | |
82 | - |
succinyl-CoA | mutant N150W, pH 7.5, 18°C | Mus musculus |