Application | Comment | Organism |
---|---|---|
analysis | development of a sensitive, non-radioactive assay for AsnS, based on incubation of desalted enzyme and substrates and then direct detection of either product asparagine or glutamate by HPLC | Zea mays |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Glycine max |
expression in Escherichia coli | Zea mays |
gene asnS, expression of C-terminally His-tagged isozyme AsnS1 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies | Zea mays |
gene asnS2, expression of nontagged isozyme ZmAsnS2 in Escherichia coli, expression of C-terminally His-tagged isozyme AsnS2 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies although small amounts of ZmAsnS2 are recovered in the soluble fraction | Zea mays |
gene asnS3, expression of C-terminally His-tagged isozyme AsnS3 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies | Zea mays |
gene asnS4, expression of C-terminally His-tagged isozyme AsnS4 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies | Zea mays |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
asparagine | - |
Glycine max | |
asparagine | competitive; competitive | Zea mays | |
glutamate | - |
Glycine max | |
glutamate | competitive; competitive | Zea mays | |
L-asparagine | competitive inhibition | Zea mays | |
L-glutamate | competitive inhibition | Zea mays |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | the AsnS isozymes are kinetically distinct with substantial differences in Km (Gln) and Vmax values, overview. None of the enzymes has cooperative enzyme kinetics | Zea mays | |
0.09 | - |
L-glutamine | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn2 | Zea mays | |
0.097 | - |
ATP | pH 7.6, 22°C | Zea mays | |
0.097 | - |
ATP | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3 | Zea mays | |
0.1 | 1 | ATP | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1 | Zea mays | |
0.1 | 1 | L-glutamine | pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2 | Zea mays | |
0.1 | 2 | L-aspartate | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3 | Zea mays | |
0.106 | - |
ATP | pH 7.6, 22°C | Glycine max | |
0.11 | - |
ATP | pH 7.6, 22°C | Zea mays | |
0.11 | - |
L-Gln | pH 7.6, 22°C | Zea mays | |
0.125 | - |
ATP | pH 7.6, 22°C | Zea mays | |
0.125 | - |
ATP | pH 7.6, temperature not specified in the publication, recombinant nontagged isozyme soluble ZmAsn2 | Zea mays | |
0.128 | - |
ATP | pH 7.6, 22°C | Zea mays | |
0.128 | - |
ATP | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn2 | Zea mays | |
0.128 | - |
ATP | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4 | Zea mays | |
0.233 | - |
L-Gln | pH 7.6, 22°C | Zea mays | |
0.233 | - |
L-glutamine | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4 | Zea mays | |
0.254 | - |
L-Gln | pH 7.6, 22°C | Glycine max | |
0.423 | - |
L-Gln | pH 7.6, 22°C | Zea mays | |
0.423 | - |
L-glutamine | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3 | Zea mays | |
0.543 | - |
L-Gln | pH 7.6, 22°C | Zea mays | |
0.543 | - |
L-glutamine | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1 | Zea mays | |
0.75 | - |
NH3 | pH 7.6, 22°C | Zea mays | |
0.85 | - |
L-Asp | pH 7.6, 22°C | Glycine max | |
0.9 | 1 | L-aspartate | pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2 | Zea mays | |
0.91 | - |
L-Asp | pH 7.6, 22°C | Zea mays | |
0.93 | - |
L-Asp | pH 7.6, 22°C | Zea mays | |
0.93 | - |
L-aspartate | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4 | Zea mays | |
0.98 | - |
L-Asp | pH 7.6, 22°C | Zea mays | |
0.98 | - |
L-aspartate | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1 | Zea mays | |
0.98 | - |
L-aspartate | pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn2 | Zea mays | |
1.2 | - |
L-Asp | pH 7.6, 22°C | Zea mays | |
8.4 | - |
NH3 | pH 7.6, 22°C | Zea mays | |
9 | - |
NH3 | pH 7.6, 22°C | Zea mays | |
9.9 | - |
NH3 | pH 7.6, 22°C | Zea mays |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Zea mays |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + L-glutamine + H2O | Zea mays | - |
AMP + diphosphate + L-asparagine + L-glutamate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Glycine max | - |
- |
- |
Zea mays | B4FFJ0 | isoform AsnS1 | - |
Zea mays | B4FFJ0 | isozyme AsnS1; isozyme AsnS1 | - |
Zea mays | B5U8J7 | isoform AsnS2 | - |
Zea mays | B5U8J7 | isozyme AsnS2; isozyme AsnS2 | - |
Zea mays | B5U8J8 | isoform AsnS3 | - |
Zea mays | B5U8J8 | isozyme AsnS3; isozyme AsnS3 | - |
Zea mays | B5U8J9 | isoform AsnS4 | - |
Zea mays | B5U8J9 | isozyme AsnS4; isozyme AsnS4 | - |
Purification (Comment) | Organism |
---|---|
- |
Glycine max |
- |
Zea mays |
development of a method to refold recombinant AsnS from inclusion bodies | Zea mays |
refolded, solubilized recombinant His-tagged isozyme AsnS1 by nickel affinity chromatography | Zea mays |
refolded, solubilized recombinant His-tagged isozyme AsnS2 by nickel affinity chromatography, recombinant nontagged isozyme ZmAsnS2 from Escherichia coli by anion exchange chromatography | Zea mays |
refolded, solubilized recombinant His-tagged isozyme AsnS4 by nickel affinity chromatography | Zea mays |
refolded, solubilized recombinant His-tagged isozyme by nickel affinity chromatography | Zea mays |
Renatured (Comment) | Organism |
---|---|
refolding of recombinant C-terminally His-tagged enzyme from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active proteins, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change | Zea mays |
refolding of recombinant C-terminally His-tagged enzymes from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active proteins, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change | Zea mays |
refolding of recombinant C-terminally His-tagged isozyme AsnS1 from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active protein, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change | Zea mays |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-Asp + L-Gln | - |
Glycine max | AMP + diphosphate + L-Asn + L-Glu | - |
? | |
ATP + L-Asp + L-Gln | - |
Zea mays | AMP + diphosphate + L-Asn + L-Glu | - |
? | |
ATP + L-Asp + NH3 | - |
Glycine max | AMP + diphosphate + L-Asn | - |
? | |
ATP + L-Asp + NH3 | - |
Zea mays | AMP + diphosphate + L-Asn | - |
? | |
ATP + L-aspartate + L-glutamine + H2O | - |
Zea mays | AMP + diphosphate + L-asparagine + L-glutamate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ASNS | - |
Zea mays |
AsnS1 | - |
Zea mays |
AsnS2 | - |
Zea mays |
AsnS3 | - |
Zea mays |
AsnS4 | - |
Zea mays |
Asparagine synthetase | - |
Zea mays |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Zea mays |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Zea mays |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.25 | - |
asparagine | pH 7.6, 22°C | Zea mays | |
0.25 | - |
L-asparagine | pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2 | Zea mays | |
0.3 | - |
glutamate | pH 7.6, 22°C | Zea mays | |
0.3 | - |
L-glutamate | pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2 | Zea mays |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
1.1 | - |
pH 7.6, 22°C | Zea mays | asparagine | |
1.1 | - |
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1 | Zea mays | L-asparagine | |
1.2 | - |
pH 7.6, 22°C | Glycine max | glutamate | |
1.2 | - |
pH 7.6, 22°C | Zea mays | glutamate | |
1.2 | - |
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4 | Zea mays | L-glutamate | |
1.3 | - |
pH 7.6, 22°C | Zea mays | glutamate | |
1.3 | - |
pH 7.6, 22°C | Glycine max | asparagine | |
1.3 | - |
pH 7.6, 22°C | Zea mays | asparagine | |
1.3 | - |
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAs4 | Zea mays | L-asparagine | |
1.3 | - |
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1 | Zea mays | L-glutamate | |
1.3 | - |
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3 | Zea mays | L-glutamate | |
1.3 | - |
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2 | Zea mays | L-glutamate | |
1.4 | - |
pH 7.6, 22°C | Zea mays | asparagine | |
1.4 | - |
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2 | Zea mays | L-asparagine | |
1.5 | - |
pH 7.6, 22°C | Zea mays | asparagine | |
1.5 | - |
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3 | Zea mays | L-asparagine |
General Information | Comment | Organism |
---|---|---|
metabolism | Asn is a major amino acid in maize and since AsnS is the primary means of Asn synthesis in plants it plays a very important role in nitrogen metabolism | Zea mays |