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Literature summary for 2.3.1.17 extracted from

  • Wang, Q.; Zhao, M.; Parungao, G.G.; Viola, R.E.
    Purification and characterization of aspartate N-acetyltransferase: a critical enzyme in brain metabolism (2016), Protein Expr. Purif., 119, 11-18.
    View publication on PubMed

Cloned(Commentary)

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 (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged truncated enzyme, X-ray diffraction structure determination and analysis Homo sapiens

Inhibitors

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 [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

Localization Comment Organism GeneOntology No. Textmining
membrane membrane-associated, the enzyme contains a membrane anchor region Homo sapiens 16020
-

Molecular Weight [Da]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens Q8N9F0
-
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining
brain
-
Homo sapiens
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms Comment Organism
ANAT
-
Homo sapiens

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Homo sapiens

pH Range

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

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