Application | Comment | Organism |
---|---|---|
additional information | upon binding to the inactive AK1-Lys complex, S-adenosyl-L-methionine promotes a slow conformational transition leading to formation of a stable aspartate kinase 1-Lys-S-adenosyl-L-methionine complex. Increase in AK1 apparent affinity for lysine in the presence of S-adenosyl-L-methionine results from the displacement of the unfavorable equilibrium between AK1 and aspartate kinase 1-Lys towards the inactive form, S-adenosyl-L-methionine and Lys binding to AK1 is sequential, with Lys binding preceding S-adenosyl-L-methionine binding | Arabidopsis thaliana |
additional information | upon binding to the inactive AK1Lys complex, S-adenosyl-l-methionine promotes a slow conformational transition leading to formation of a stable aspartate kinase 1LysS-adenosyl-l-methionine complex. Increase in AK1 apparent affinity for lysine in the presence of S-adenosyl-l-methionine results from the displacement of the unfavorable equilibrium between AK1 and aspartate kinase 1Lys towards the inactive form, S-adenosyl-l-methionine and Lys binding to AK1 is sequential, with Lys binding preceding S-adenosyl-l-methionine binding | Arabidopsis thaliana |
Cloned (Comment) | Organism |
---|---|
ligated into pET 23d(+) vector and Escherichia coli strain DH10B for cloning, expression in Escherichia coli BL21 (DE3) pLysS codon+ | Arabidopsis thaliana |
ligated into pET 23d(+) vector, expressed in Escherichia coli BL21 codon (+) | Arabidopsis thaliana |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-lysine | AK1 is inhibited in a synergistic manner by lysine and S-adenosyl-L-methionine, in the absence of S-adenosyl-L-methionine AK1 displays low apparent affinity for lysine compared to AK2 and AK3; AK1 is inhibited in a synergistic manner by lysine and SAM; AK2 is inhibited by lysine, SAM by itself has no effect on the enzyme activity; AK3 is inhibited only by lysine, SAM by itself has no effect on the enzyme activity | Arabidopsis thaliana | |
additional information | no additional inhibition is observed upon addition of Thr or Leu in the presence of 100 microM Lys and 20 microM SAM; no additional inhibition is observed upon addition of Thr or Leu in the presence of 5 microM Lys for AK2; no additional inhibition is observed upon addition of Thr or Leu in the presence of 5 microM Lys for AK3; the other amino acids tested (Met, Gln, Asn, Glu, Arg) have no effect on the enzyme activities at 2.5 mM either in the presence or the absence of the inhibitor Lys or Lys plus SAM | Arabidopsis thaliana | |
S-adenosyl-L-methionine | AK1 is inhibited in a synergistic manner by lysine and S-adenosyl-L-methionine, in the presence of S-adenosyl-L-methionine, the apparent affinity of AK1 for lysine increases considerably for lysine inhibition similar to those of AK2 and AK3 | Arabidopsis thaliana |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.56 | - |
ATP | - |
Arabidopsis thaliana | |
0.56 | - |
ATP | AK3, 30°C, 50 mM HEPES, pH 8.0, 150 mM KCl, 20 mM MgCl2 with a coupled assay using aspartate semialdehyde dehydrogenase from Arabidopsis thaliana | Arabidopsis thaliana | |
0.98 | - |
ATP | AK2, 30°C, 50 mM HEPES, pH 8.0, 150 mM KCl, 20 mM MgCl2 with a coupled assay using aspartate semialdehyde dehydrogenase from Arabidopsis thaliana | Arabidopsis thaliana | |
1.095 | - |
L-aspartate | - |
Arabidopsis thaliana | |
1.095 | - |
aspartate | AK3, 30°C, 50 mM HEPES, pH 8.0, 150 mM KCl, 20 mM MgCl2 with a coupled assay using aspartate semialdehyde dehydrogenase from Arabidopsis thaliana | Arabidopsis thaliana | |
1.7 | - |
ATP | - |
Arabidopsis thaliana | |
1.7 | - |
ATP | 30°C, 50 mM HEPES, pH 8.0, 150 mM KCl, 20 mM MgCl2 with a coupled assay using aspartate semialdehyde dehydrogenase from Arabidopsis thaliana | Arabidopsis thaliana | |
1.94 | - |
aspartate | AK2, 30°C, 50 mM HEPES, pH 8.0, 150 mM KCl, 20 mM MgCl2 with a coupled assay using aspartate semialdehyde dehydrogenase from Arabidopsis thaliana | Arabidopsis thaliana | |
2.037 | - |
L-aspartate | - |
Arabidopsis thaliana | |
2.037 | - |
aspartate | 30°C, 50 mM HEPES, pH 8.0, 150 mM KCl, 20 mM MgCl2 with a coupled assay using aspartate semialdehyde dehydrogenase from Arabidopsis thaliana | Arabidopsis thaliana |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Arabidopsis thaliana | 9507 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
53200 | - |
gel filtration | Arabidopsis thaliana |
53200 | - |
monofunctional AK2, proteins are separated on a 10% polyacrylamide (w/v) slab gel under denaturing conditions and stained with Coomassie brilliant blue R-250 | Arabidopsis thaliana |
55100 | - |
gel filtration | Arabidopsis thaliana |
55100 | - |
monofunctional AK3, proteins are separated on a 10% polyacrylamide (w/v) slab gel under denaturing conditions and stained with Coomassie brilliant blue R-250 | Arabidopsis thaliana |
55900 | - |
gel filtration | Arabidopsis thaliana |
55900 | - |
monofunctional AK1, proteins are separated on a 10% polyacrylamide (w/v) slab gel under denaturing conditions and stained with Coomassie brilliant blue R-250 | Arabidopsis thaliana |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate | Arabidopsis thaliana | - |
ADP + 4-phospho-L-aspartate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | - |
- |
- |
Arabidopsis thaliana | O23653 | ligated into pET 23d(+) vector and Escherichia coli strain DH10B for cloning, expression in Escherichia coli BL21 (DE3) pLysS codon+ | - |
Arabidopsis thaliana | Q9LYU8 | - |
- |
Arabidopsis thaliana | Q9LYU8 | ligated into pET 23d(+) vector and Escherichia coli strain DH10B for cloning, expression in Escherichia coli BL21 (DE3) pLysS codon+ | - |
Arabidopsis thaliana | Q9S702 | - |
- |
Arabidopsis thaliana | Q9S702 | ligated into pET 23d(+) vector and Escherichia coli strain DH10B for cloning, expression in Escherichia coli BL21 (DE3) pLysS codon+ | - |
Purification (Comment) | Organism |
---|---|
on anion exchange column and by gel filtration | Arabidopsis thaliana |
weak anion exchange chromatography, affinity chromatogrphy for AK1 | Arabidopsis thaliana |
weak anion exchange chromatography, gel filtration | Arabidopsis thaliana |
Storage Stability | Organism |
---|---|
-80°C, HEPES buffer, pH 7.5, 1 mM Lys, 10% (v/v) glycerol, several month, without any loss of activity | Arabidopsis thaliana |
-80°C, HEPES buffer, pH 7.5, 1 mm Lys, 10% (v/v) glycerol, several months | Arabidopsis thaliana |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate | - |
Arabidopsis thaliana | ADP + 4-phospho-L-aspartate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AK1 | - |
Arabidopsis thaliana |
AK2 | - |
Arabidopsis thaliana |
AK3 | - |
Arabidopsis thaliana |
aspartate kinase | - |
Arabidopsis thaliana |
aspartate kinase 1 | - |
Arabidopsis thaliana |
aspartate kinase 2 | - |
Arabidopsis thaliana |
aspartate kinase 3 | - |
Arabidopsis thaliana |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
AK3 is highly unstable when stored at room temperature, losing 95% of its activity in 24 h | Arabidopsis thaliana |
additional information | - |
AK1 retaines all activity when stored at room temperature | Arabidopsis thaliana |
additional information | - |
AK2 retaines all activity when stored at room temperature | Arabidopsis thaliana |
additional information | - |
AK3 proves to be highly unstable when stored at room temperature, losing 95% of its activity in 24 h | Arabidopsis thaliana |
20 | - |
AK3 is highly unstable when stored at room temperature, losing 95% of its activity in 24 h. During this period of time, AK1 and AK2 retained all their activity | Arabidopsis thaliana |
20 | - |
in 24 h, AK1 retains all its activity | Arabidopsis thaliana |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8.4 | - |
ATP | - |
Arabidopsis thaliana | |
8.4 | - |
ATP | monofunctional AK3 | Arabidopsis thaliana | |
14.5 | - |
ATP | - |
Arabidopsis thaliana | |
14.5 | - |
ATP | monofunctional AK2 | Arabidopsis thaliana | |
23.4 | - |
ATP | - |
Arabidopsis thaliana | |
23.4 | - |
ATP | monofunctional AK1 | Arabidopsis thaliana |