Application | Comment | Organism |
---|---|---|
pharmacology | arylamine N-acetyltransferases have a key role in the detoxication and metabolic activation of numerous xenobiotics, including therapeutic drugs and carcinogens | Mesorhizobium loti |
Cloned (Comment) | Organism |
---|---|
expressed as a His-tagged fusion protein | Mesorhizobium loti |
expressed in Escherichia as a His-tagged protein | Mesorhizobium loti |
into the pET28a vector for expression in Escherichia coli cells | Mesorhizobium loti |
Protein Variants | Comment | Organism |
---|---|---|
G129A | final protein yield similar to wild-type: approximately 30 mg. Enzymatic activity: 4.758 micromol/min/mg, Km (mM): 0.184 (substrate: p-aminosalicylic acid) | Mesorhizobium loti |
G129A | mutation of the conserved glycine residue in the active-site P-loop | Mesorhizobium loti |
G129A | protein production similar to wild-type | Mesorhizobium loti |
G129I | mutation of the conserved glycine residue in the active-site P-loop | Mesorhizobium loti |
G129I | only low levels of protein production, NAT activity: 0.034 micromol/min/mg | Mesorhizobium loti |
G129I | only small amount of NAT protein, final protein yield: 0.75-0.5 mg of protein. Enzymatic activity: very low, not accurately measurable | Mesorhizobium loti |
G129P | mutation of the conserved glycine residue in the active-site P-loop | Mesorhizobium loti |
G129P | only low levels of protein production, NAT activity: 0.023 micromol/min/mg | Mesorhizobium loti |
G129P | only small amount of NAT protein, final protein yield: 0.75-0.5 mg of protein. Enzymatic activity: very low, not accurately measurable | Mesorhizobium loti |
G129S | final protein yield similar to wild-type: approximately 30 mg. Enzymatic activity: 4.152 micromol/min/mg, Km (mM): 0.447 (substrate: p-aminosalicylic acid) | Mesorhizobium loti |
G129S | mutation of the conserved glycine residue in the active-site P-loop | Mesorhizobium loti |
G129S | protein production similar to wild-type | Mesorhizobium loti |
G129T | mutation of the conserved glycine residue in the active-site P-loop | Mesorhizobium loti |
G129T | only low levels of protein production, NAT activity: 4.758 micromol/min/mg | Mesorhizobium loti |
G129T | only small amount of NAT protein, final protein yield: 0.75-0.5 mg of protein. Enzymatic activity: 0.034 micromol/min/mg | Mesorhizobium loti |
G129V | mutation of the conserved glycine residue in the active-site P-loop | Mesorhizobium loti |
G129V | only low levels of protein production, NAT activity: 4.152 micromol/min/mg | Mesorhizobium loti |
G129V | only small amount of NAT protein, final protein yield: 0.75-0.5 mg of protein. Enzymatic activity: 0.022 micromol/min/mg | Mesorhizobium loti |
additional information | it is shown that a small amino acid (such as Gly or Ala) at position 429 (start of the active-site P-loop) is important not only for maintaining the functions of the active-site P-loop but also for maintaining the overall structural integrity of NAT enzymes. Results suggest that in addition to its role in substrate binding and selectivity, the active-site P-loop plays a wider structural role in NAT enzymes | Mesorhizobium loti |
additional information | results show that amino acid 129, which is the start of the active-site P-loop, is important to be a small amino acid (such as Gly or Ala) not only for maintaining the functions of the active-site P-loop but also for maintaining the overall structural integrity of NAT enzymes. Results suggest that in addition to its role in substrate binding and selectivity, the active-site P-loop plays a wider structural role in NAT enzymes | Mesorhizobium loti |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.184 | - |
p-Aminosalicylic acid | mutant G129A | Mesorhizobium loti | |
0.184 | - |
PAS | mutant G129A | Mesorhizobium loti | |
0.216 | - |
p-Aminosalicylic acid | - |
Mesorhizobium loti | |
0.216 | - |
PAS | wild-type | Mesorhizobium loti | |
0.447 | - |
p-Aminosalicylic acid | mutant G129S | Mesorhizobium loti | |
0.447 | - |
PAS | mutant G129S | Mesorhizobium loti |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34000 | - |
determined by SDS-PAGE and Western Blot analysis | Mesorhizobium loti |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mesorhizobium loti | - |
- |
- |
Purification (Comment) | Organism |
---|---|
using Ni-NTA chromatography. Protein yield: approximately 30 mg | Mesorhizobium loti |
using Ni2+-agarose | Mesorhizobium loti |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
enzymatic activity of mutants G129I and G129P are very low and not accurately measurable | Mesorhizobium loti |
additional information | - |
mutant G129I and G129P display only very low enzymatic activity which can not be measured accurately | Mesorhizobium loti |
0.022 | - |
mutant G129V | Mesorhizobium loti |
0.023 | - |
mutant G129V | Mesorhizobium loti |
0.034 | - |
mutant G129T | Mesorhizobium loti |
4.152 | - |
mutant G129S | Mesorhizobium loti |
4.758 | - |
mutant G129A | Mesorhizobium loti |
5.078 | - |
wild-type enzyme | Mesorhizobium loti |
5.078 | - |
wild-type | Mesorhizobium loti |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 2-aminofluorene | - |
Mesorhizobium loti | CoA + N-acetyl-2-aminofluorene | - |
? | |
acetyl-CoA + p-aminosalicylate | - |
Mesorhizobium loti | CoA + N-acetyl-p-aminosalicylate | - |
? | |
acetyl-CoA + p-aminosalicylic acid | - |
Mesorhizobium loti | CoA + N-acetyl-4-aminosalicylic acid | - |
? |
Synonyms | Comment | Organism |
---|---|---|
arylamine N-acetyltransferase 1 | - |
Mesorhizobium loti |
MlNAT1 | - |
Mesorhizobium loti |
NAT | - |
Mesorhizobium loti |
NAT1 | - |
Mesorhizobium loti |