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Literature summary extracted from

  • Hurtado-Guerrero, R.; Raimi, O.G.; Min, J.; Zeng, H.; Vallius, L.; Shepherd, S.; Ibrahim, A.F.; Wu, H.; Plotnikov, A.N.; van Aalten, D.M.
    Structural and kinetic differences between human and Aspergillus fumigatus D-glucosamine-6-phosphate N-acetyltransferase (2008), Biochem. J., 415, 217-223.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.4 expressed in Escherichia coli as a GST-fusion protein Aspergillus fumigatus
2.3.1.4 expressed in Escherichia coli as a His-tagged-fusion protein Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.1.4 crystal structures of human and Aspergillus fumigatus GNA1 are determined: structural differences between the two enzymes are mostly located to the sugar-binding site, whereas the AcCoA-binding site is more conserved. These changes affect not only the electrostatics, but also reveal a more spacious sugar binding site in the Aspergillus fumigatus GNA1 enzyme, whereas large side chains at these positions create a tighter pocket in the HsGNA1 enzyme Aspergillus fumigatus
2.3.1.4 crystal structures of human and Aspergillus fumigatus GNA1 are determined: structural differences between the two enzymes are mostly located to the sugar-binding site, whereas the AcCoA-binding site is more conserved. These changes affect not only the electrostatics, but also reveal a more spacious sugar binding site in the Aspergillus fumigatus GNA1 enzyme, whereas large side chains at these positions create a tighter pocket in the HsGNA1 enzyme Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.4 E185Y Km (mM): 0.03 (D-glucosamine 6-phosphate), 0.04 (acetyl-CoA), kcat (1/sec): 1.34 (D-glucosamine 6-phosphate) Aspergillus fumigatus
2.3.1.4 G183E Km (mM): 0.056 (D-glucosamine 6-phosphate), 0.1 (acetyl-CoA), kcat (1/sec): 0.34 (D-glucosamine 6-phosphate) Aspergillus fumigatus
2.3.1.4 additional information mutation of the four nonconserved residues (V125, G183, E185 and Y189) lining the sugar-binding pocket suggests that they contribute to the kinetic differences between GNA1 from Aspergillus fumigatus and GNA1 from Homo sapiens Aspergillus fumigatus
2.3.1.4 V125R Km (mM): 0.6 (D-glucosamine 6-phosphate), 0.2 (acetyl-CoA), kcat (1/sec): 0.12 (D-glucosamine 6-phosphate) Aspergillus fumigatus
2.3.1.4 Y189R Km (mM): 0.056 (D-glucosamine 6-phosphate), 0.086 (acetyl-CoA), kcat (1/sec): 1.7 (D-glucosamine 6-phosphate) Aspergillus fumigatus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.4 0.026
-
 acetyl-CoA wild-type Homo sapiens
2.3.1.4 0.03
-
 D-glucosamine 6-phosphate mutant E185Y Aspergillus fumigatus
2.3.1.4 0.04
-
 acetyl-CoA wild-type Aspergillus fumigatus
2.3.1.4 0.04
-
 acetyl-CoA mutant E185Y Aspergillus fumigatus
2.3.1.4 0.056
-
 D-glucosamine 6-phosphate mutant G183E Aspergillus fumigatus
2.3.1.4 0.056
-
 D-glucosamine 6-phosphate mutant Y189R Aspergillus fumigatus
2.3.1.4 0.071
-
 D-glucosamine 6-phosphate wild-type Aspergillus fumigatus
2.3.1.4 0.086
-
 acetyl-CoA mutant Y189R Aspergillus fumigatus
2.3.1.4 0.097
-
 D-glucosamine 6-phosphate wild-type Homo sapiens
2.3.1.4 0.1
-
 acetyl-CoA mutant G183E Aspergillus fumigatus
2.3.1.4 0.2
-
 acetyl-CoA mutant V125R Aspergillus fumigatus
2.3.1.4 0.6
-
 D-glucosamine 6-phosphate mutant V125R Aspergillus fumigatus

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.4 Aspergillus fumigatus
-
-
-
2.3.1.4 Homo sapiens Q96EK6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.4 using affinity, ion-exchange and size-exclusion chromatography: yield 25 mg of pure HsGNA1 per litre of bacterial culture Homo sapiens
2.3.1.4 using affinity, ion-exchange and size-exclusion chromatography: yield 4 mg of pure AfGNA1 per litre of bacterial culture Aspergillus fumigatus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.1.4 25
-
-
 Homo sapiens
2.3.1.4 107
-
-
 Aspergillus fumigatus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.4 acetyl-CoA + D-glucosamine 6-phosphate
-
 Aspergillus fumigatus CoA + N-acetyl-D-glucosamine 6-phosphate
-
 ?
2.3.1.4 acetyl-CoA + D-glucosamine 6-phosphate
-
 Homo sapiens CoA + N-acetyl-D-glucosamine 6-phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.1.4 homodimer crystal structure Aspergillus fumigatus
2.3.1.4 homodimer crystal structure Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
2.3.1.4 AfGNA1
-
 Aspergillus fumigatus
2.3.1.4 glucosamine-6-phosphate N-acetyltransferase
-
 Aspergillus fumigatus
2.3.1.4 glucosamine-6-phosphate N-acetyltransferase
-
 Homo sapiens
2.3.1.4 HsGNA1
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.4 additional information
-
 assay carried out at room temperature Aspergillus fumigatus
2.3.1.4 additional information
-
 assay carried out at room temperature Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.4 0.12
-
 D-glucosamine 6-phosphate mutant V125R Aspergillus fumigatus
2.3.1.4 0.34
-
 D-glucosamine 6-phosphate mutant G183E Aspergillus fumigatus
2.3.1.4 1.34
-
 D-glucosamine 6-phosphate mutant E185Y Aspergillus fumigatus
2.3.1.4 1.7
-
 D-glucosamine 6-phosphate mutant Y189R Aspergillus fumigatus
2.3.1.4 9
-
 D-glucosamine 6-phosphate wild-type Homo sapiens
2.3.1.4 38
-
 D-glucosamine 6-phosphate wild-type Aspergillus fumigatus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.4 7.5
-
 assay at Aspergillus fumigatus
2.3.1.4 7.5
-
 assay at Homo sapiens