Crystallization (Comment) | Organism |
---|---|
crystal structure of the Glu317Gln mutant in complex with UDP-Gal is reported | Bos taurus |
purified recombinant enzyme mutants with bound UDP-Gal and Mn2+, hanging drop vapour diffusion at 16°C, mixing of 0.001 ml protein solution containing 5 mg/ml mutant E317Q in 20 mM MES-NaOH buffer, pH 6.0, and 10% glycerol, containing 10 mM MnCl2 and 10 mM UDP-Gal with 0.001 ml of reservoir solution containing 10% PEG 6000, 0.1 M Tris-HCl, pH 8.0, and 8% MPD, for mutant D316N and D316E, 0.001 ml of 5 mg/ml protein in in 20 mM MES-NaOH buffer, pH 6.0, with 10% glycerol, 10 mM UDP-Gal, 10 mM MnCl2, and 10 mM N-acetyllactosamine, are mixed with 0.001 ml of a reservoir solution containing 5-10% PEG 6000, 0.1 M Tris-HCl, pH 8.0, and 5-14% MPD, X-ray diffraction structure determination and analysis at 1.77-2.2 A resolution | Bos taurus |
Protein Variants | Comment | Organism |
---|---|---|
D316E | mutant show modest reduction in kcat and Km for lactose. Strucutural studies with mutant D316E show that the negative charge is crucial for catalytic activity and needed for its interaction with Arg202 for an active site structure that facilitates the binding of UDP-gal in a catalytically competent conformation | Bos taurus |
D316E | site-directed mutagenesis, a catalytic domain mutant, crystal structure determination with bound UDP-Gal and Mn2+ | Bos taurus |
D316N | mutant is inactive. Strucutural studies with mutant D316N show that the negative charge is crucial for catalytic activity and needed for its interaction with Arg202 for an active site structure that facilitates the binding of UDP-gal in a catalytically competent conformation | Bos taurus |
D316N | site-directed mutagenesis, a catalytic domain mutant, crystal structure determination with bound UDP-Gal and Mn2+ | Bos taurus |
E317Q | crystal structure of the complex of mutant E317Q with UDP-galactose exhibiting a bent configuration stabilized by interactions of the galactose with multiple residues in the enzyme including those in a highly conserved region (His315 to Ser318) is shown | Bos taurus |
E317Q | site-directed mutagenesis, a catalytic domain mutant, crystal structure determination with bound UDP-Gal and Mn2+ | Bos taurus |
H315Q | mutant protein shows modest changes in kinetic parameters for lactose | Bos taurus |
H315R | mutant shows a major losss in catalytic activity arising from a 500fold reduction in kcat | Bos taurus |
H319A | mutant shows a 2fold reduction for kcat | Bos taurus |
H319E | mutant is inactive | Bos taurus |
H319Y | mutant is inactive | Bos taurus |
S318A | mutant shows a 10fold reduction for kcat | Bos taurus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics | Bos taurus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | binding structure at the catalytic domain, structure model, overview | Bos taurus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bos taurus | - |
- |
- |
Bos taurus | P14769 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | structural basis of UDP-galactose binding at the catalytic domain of alpha3GT, overview | Bos taurus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
alpha-1,3-galactosyltransferase | - |
Bos taurus |
alpha3GT | - |
Bos taurus |