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Literature summary for 2.3.1.5 extracted from

  • Atmane, N.; Dairou, J.; Flatters, D.; Martins, M.; Pluvinage, B.; Derreumaux, P.; Dupret, J.M.; Rodrigues-Lima, F.
    The conserved glycine/alanine residue of the active-site loop containing the putative acetylCoA-binding motif is essential for the overall structural integrity of Mesorhizobium loti arylamine N-acetyltransferase 1 (2007), Biochem. Biophys. Res. Commun., 361, 256-262.
    View publication on PubMed

Application

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(Commentary)

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

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
determined by SDS-PAGE and Western Blot analysis Mesorhizobium loti

Organism

Organism UniProt Comment Textmining
Mesorhizobium loti
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography. Protein yield: approximately 30 mg Mesorhizobium loti
using Ni2+-agarose Mesorhizobium loti

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
arylamine N-acetyltransferase 1
-
Mesorhizobium loti
MlNAT1
-
Mesorhizobium loti
NAT
-
Mesorhizobium loti
NAT1
-
Mesorhizobium loti