Cloned (Comment) | Organism |
---|---|
gene nat8l, sequence comparisons, subcloning of the genes for thioredoxin (TRX), glutathione S-transferase (GST) or maltose binding protein (MBP), followed by a linker region (21-68-amino acids) containing various cleavage site sequences, and then connected to the N-terminal of the nat8l gene, without the membrane anchor, recombinant functional and soluble expression of the dual affinity tagged enzyme as MBP-fusion protein in Escherichia coli strain BL21(DE3), method evaluation | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged truncated enzyme, X-ray diffraction structure determination and analysis | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
cholamido propane sulfonate | - |
Homo sapiens | |
cyclohexylmaltoside | cymal5, high inhibition at CMC concentration | Homo sapiens | |
decylmaltoside | - |
Homo sapiens | |
DMSO | 20% inhibition at 40% v/v | Homo sapiens | |
dodecyl octaglycol | - |
Homo sapiens | |
dodecylmaltoside | - |
Homo sapiens | |
lauryldimethylamine-N-oxide | complete inhibition at CMC concentration | Homo sapiens | |
additional information | effect of different detergents on the enzyme activity: non-ionic detergents such as Triton X-100 are less disruptive to protein structures than ionic detergents such as SDS, detergents such as C12E8, Tween 20 and several maltosides caused minimal disruption of the enzyme, with greater than 50% residual activity after incubation with CMC levels of each of these detergents. In contrast, significant loss of activity is observed upon incubation with C8 detergents, cymal5, octylglucoside and some shorter chain polymaleic anhydride (pmal) detergents. Ionic detergent SDS and a zwitterionic detergent lauryldimethylamine-N-oxide cause nearly complete loss of catalytic activity | Homo sapiens | |
n-decyl-N,N-dimethylamine-N-oxide | high inhibition at CMC concentration | Homo sapiens | |
N-methyl-N-nonanoyl-beta-D-glucosylamine | Mega-9, high inhibition at CMC concentration | Homo sapiens | |
n-nonyl-beta-D-glucopyranoside | high inhibition at CMC concentration | Homo sapiens | |
octyl pentaglycol | high inhibition at CMC concentration | Homo sapiens | |
octyl tetraglycol | high inhibition at CMC concentration | Homo sapiens | |
octylglucoside | high inhibition at CMC concentration | Homo sapiens | |
polymaleic anhydride C10 | - |
Homo sapiens | |
polymaleic anhydride C12 | - |
Homo sapiens | |
polymaleic anhydride C16 | high inhibition at CMC concentration | Homo sapiens | |
polymaleic anhydride C4 | high inhibition at CMC concentration | Homo sapiens | |
polymaleic anhydride C6 | complete inhibition at CMC concentration | Homo sapiens | |
polymaleic anhydride C8 | - |
Homo sapiens | |
SDS | complete inhibition at CMC concentration | Homo sapiens | |
sodium dodecanoyl sarcosine | - |
Homo sapiens | |
Triton X-100 | - |
Homo sapiens | |
Tween 20 | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Homo sapiens | |
0.0031 | - |
acetyl-CoA | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.16 | - |
L-aspartate | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.36 | - |
3-methyl-L-aspartate | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.92 | - |
2,3-diaminosuccinate | pH 7.4, temperature not specified in the publication | Homo sapiens | |
8.6 | - |
L-glutamate | pH 7.4, temperature not specified in the publication | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | membrane-associated, the enzyme contains a membrane anchor region | Homo sapiens | 16020 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
33900 | - |
x * 56000-60000, recombinant MBP-His-tagged enzyme without membrane anchor, SDS-PAGE, x * 33900, recombinant His-tagged enzyme without membrane anchor, SDS-PAGE | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + L-aspartate | Homo sapiens | - |
CoA + N-acetyl-L-aspartate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q8N9F0 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant functional and soluble dual His- and MBP-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and amylose affinity chromatography, MBP tag cleavage by 3C protease, method evaluation | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
brain | - |
Homo sapiens | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
70 | - |
recombinant enzyme, pH 7.4, temperature not specified in the publication | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 2,3-diaminosuccinate | - |
Homo sapiens | CoA + ? | - |
? | |
acetyl-CoA + 3-methyl-L-aspartate | - |
Homo sapiens | CoA + N-acetyl-3-methyl-L-aspartate | - |
? | |
acetyl-CoA + L-aspartate | - |
Homo sapiens | CoA + N-acetyl-L-aspartate | - |
? | |
acetyl-CoA + L-glutamate | reaction of EC 2.3.1.1 | Homo sapiens | CoA + N-acetyl-L-glutamate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 56000-60000, recombinant MBP-His-tagged enzyme without membrane anchor, SDS-PAGE, x * 33900, recombinant His-tagged enzyme without membrane anchor, SDS-PAGE | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
ANAT | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0018 | - |
3-methyl-L-aspartate | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.0071 | - |
acetyl-CoA | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.0071 | - |
L-aspartate | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.023 | - |
L-glutamate | pH 7.4, temperature not specified in the publication | Homo sapiens | |
0.035 | - |
2,3-diaminosuccinate | pH 7.4, temperature not specified in the publication | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Homo sapiens |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 9.5 | the enzymatic activity decreases at pH values below pH 7.5. A fit of the Vmax/Km data to a model which assumes that the protonation of a single group leads to loss of activity results in a pK value of 6.8 for a group that must be ionized for the enzyme to remain catalytically active. By contrast, the Vmax profile does not show substantial changes across the pH range | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | Canavan disease is a fatal, neurological disease that is caused by an interruption in the metabolism of a critical amino acid, N-acetyl-L-aspartic acid. Defects at multiple locations in the aspA gene that codes for aspartoacylase, EC 3.5.1.15, lead to mutant forms of this enzyme that are either not expressed or rapidly degraded, or have significantly impaired catalytic activity, resulting in N-acetyl-L-aspartic acid accumulation. A second gene knock-out in the Nat8l gene which codes for aspartate N-acetyltransferase, the enzyme that synthesizes N-acetyl-L-aspartic acid, reverses these adverse effects, leading to normal myelination and a decrease in Canavan disease symptoms | Homo sapiens |