Application | Comment | Organism |
---|---|---|
synthesis | production of novel complex glycans with role in biological recognition processes with engineered enzyme with modified substrate specificity | Bos taurus |
Cloned (Comment) | Organism |
---|---|
PCR megaprimer method, expressed in Escherichia coli BL21(DE3) | Bos taurus |
Crystallization (Comment) | Organism |
---|---|
AGGL-mutant in complex with UDP-N-acetylgalactosamine with 20 mM MES-NaOH buffer, pH 6, 10% glycerol, 10 mM MnCl2, 10 mM UDP-N-acetylgalactosamine and reservoir solution (10-15% PEG 6000, 0.1 M Tris-HCl, pH 8, 15-25% MPD), and RAAI-mutant with 0.1 M Tris-HCl, pH 8, 10-50% PEG 4000, 0.2 M sodium acetate, 10 mM MnCl2, 10 mM UDP-galactose with the same reservoir solution, vapor diffusion hanging drop method | Bos taurus |
Protein Variants | Comment | Organism |
---|---|---|
H280A/A281G/A282G/L283I | AGGI-mutant, instable, leucin 283 is important for enzyme stability | Bos taurus |
H280A/A281G/A282G/L283L | AGGL-mutant, higher N-acetylgalactosamine transferase activity than galactosyltransferase activity, leucin 283 is important for enzyme stability | Bos taurus |
H280A/A281G/A282G/L283V | AGGV-mutant, instable, leucin 283 is important for enzyme stability | Bos taurus |
H280G/A281G/A282G/L283I | GGGI-mutant, instable, leucin 283 is important for enzyme stability | Bos taurus |
H280G/A281G/A282G/L283L | GGGL-mutant, higher N-acetylgalactosamine transferase activity than galactosyltransferase activity, leucin 283 is important for enzyme stability | Bos taurus |
H280G/A281G/A282G/L283V | GGGV-mutant, instable, leucin 283 is important for enzyme stability | Bos taurus |
H280R/A281A/A282A/L283I | RAAI-mutant, no catalytic activity, crystallisation | Bos taurus |
additional information | mutants with modified donor substrate specificity | Bos taurus |
General Stability | Organism |
---|---|
mutants are not stable under assay conditions (10 mM Tris-HCl, pH 7.4, 37°C), but for 20 min with 10 mM MES, pH 6.0, at 30°C | Bos taurus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
UDP-galactose | AGGL mutant, lactose, pH 6, 30°C | Bos taurus | |
0.12 | - |
UDP-galactose | wild-type, N-acetyllactosamine, pH 6, 30°C | Bos taurus | |
0.15 | - |
UDP-galactose | AGGL mutant, N-acetyllactosamine, pH 6, 30°C | Bos taurus | |
0.16 | - |
UDP-galactose | wild-type, lactose, pH 6, 30°C | Bos taurus | |
0.25 | - |
UDP-N-acetylgalactosamine | AGGL mutant, lactose, pH 6, 30°C | Bos taurus | |
0.6 | - |
N-acetyllactosamine | wild-type, UDP-galactose, pH 6, 30°C | Bos taurus | |
0.64 | - |
UDP-N-acetylgalactosamine | AGGL mutant, N-acetyllactosamine, pH 6, 30°C | Bos taurus | |
13 | - |
N-acetyllactosamine | AGGL mutant, UDP-galactose, pH 6, 30°C | Bos taurus | |
15 | - |
N-acetyllactosamine | AGGL mutant, UDP-N-acetylgalactosamine, pH 6, 30°C | Bos taurus | |
30 | - |
lactose | wild-type, UDP-galactose, pH 6, 30°C | Bos taurus | |
98 | - |
lactose | AGGL mutant, UDP-N-acetylgalactosamine, pH 6, 30°C | Bos taurus | |
100 | - |
lactose | AGGL mutant, UDP-galactose, pH 6, 30°C | Bos taurus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
MnCl2 | - |
Bos taurus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-galactose + N-acetyllactosamine | Bos taurus | - |
UDP + ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bos taurus | P14769 | - |
- |
Purification (Comment) | Organism |
---|---|
centrifugation of cell culture, pellet lysed with EasyLyse, loaded onto Ni-chelate column, washing and elution with 20 mM Tris-HCl, pH 7.9, containing NaCl and imidazole | Bos taurus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-galactose + N-acetyllactosamine | - |
Bos taurus | UDP + ? | - |
? | |
UDP-galactose + lactose | - |
Bos taurus | UDP + ? | - |
? | |
UDP-galactose + N-acetyllactosamine | - |
Bos taurus | UDP + ? | - |
? | |
UDP-N-acetylgalactosamine + lactose | no reaction of wild-type, only of AGGL mutant | Bos taurus | UDP + ? | - |
? | |
UDP-N-acetylgalactosamine + N-acetyllactosamine | no reaction of wild-type, only of AGGL mutant | Bos taurus | UDP + ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
alpha-1,3-galactosyltransferase | - |
Bos taurus |
alpha3GT | - |
Bos taurus |
UDP-galactose beta-galactosyl alpha-1,3-galactosyltransferase | - |
Bos taurus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0067 | - |
UDP-galactose | AGGL mutant, N-acetyllactosamine, pH 6, 30°C | Bos taurus | |
0.0099 | - |
UDP-galactose | AGGL mutant, lactose, pH 6, 30°C | Bos taurus | |
0.045 | - |
UDP-N-acetylgalactosamine | AGGL mutant, N-acetyllactosamine, pH 6, 30°C | Bos taurus | |
0.095 | - |
UDP-N-acetylgalactosamine | AGGL mutant, lactose, pH 6, 30°C | Bos taurus | |
1.91 | - |
UDP-galactose | wild-type, N-acetyllactosamine, pH 6, 30°C | Bos taurus | |
4.8 | - |
UDP-galactose | wild-type, lactose, pH 6, 30°C | Bos taurus |