Crystallization (Comment) | Organism |
---|---|
M344H mutant enzyme complexed with UDP-Gal and Mn2+ or Mg2+, hanging drop vapour diffusion method, 30 mg/ml protein in solution with 17 mM UDP-Gal, and 17 mM MgCl2 or MnCl2, precipitated by 10% v/v dioxane, 100 mM MES-NaOH, pH 6.5, and 2.0M ammonium sulfate, room temperature, X-ray diffraction structure determination and analysis at 2.3 A resolution, determination of conformation stereochemistry, modeling | Bos taurus |
Protein Variants | Comment | Organism |
---|---|---|
M344A | site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme | Bos taurus |
M344E | site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme | Bos taurus |
M344H | site-directed mutagenesis, the mutant shows altered conformational changes upon binding of Mn2+ and substrates or substrate analogues compared to the wild-type enzyme, it forms the closed conformation with bound Mn2+ and UDP-hexanolamine, loss of 98% of Mn2+ binding activity, increased activity with Mg2+ | Bos taurus |
M344Q | site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme | Bos taurus |
M344S | site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme | Bos taurus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics, wild-type and mutant M344H enzymes | Bos taurus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
Golgi apparatus | - |
Bos taurus | 5794 | - |
membrane | type II membrane-bound protein | Bos taurus | 16020 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | active only with mutant enzymes M334H and M344E, no activity with wild-type enzyme and mutants M344A, M344S, and M344Q | Bos taurus | |
Co2+ | less active than Mn2+ with wild-type and mutant enzymes | Bos taurus | |
Mg2+ | active only with mutant enzymes M334H and M344E, no activity with wild-type enzyme and mutants M344A, M344S, and M344Q | Bos taurus | |
Mn2+ | required, metal binding site structure with an uncommon coordination to the Sdelta atom of Met344, the site is located in the hinge region of along flexible loop, which changes from open to closed conformation upon binding of Mn2+ and UDP-galactose creating an oligosaccharide binding site in the enzyme structure, effect occurs specifically with UDP-galactose, not with other UDP-sugars,overview | Bos taurus | |
additional information | metal ion specificity of wild-type and mutant enzymes, overview | Bos taurus | |
Sr2+ | active only with mutant enzymes M334H and M344E, no activity with wild-type enzyme and mutants M344A, M344S, and M344Q | Bos taurus | |
Zn2+ | less active than Mn2+ with wild-type and mutant enzymes | Bos taurus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-galactose + N-acetyl-D-glucosamine | Bos taurus | - |
UDP + N-acetyllactosamine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bos taurus | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
UDP-alpha-D-galactose + N-acetyl-D-glucosamine = UDP + N-acetyllactosamine | the GlcNAc residue at the nonreducing end of chitobiose makes extensive hydrophobic interactions with the highly conserved Tyr286 | Bos taurus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
metal ion specificity of wild-type and mutant enzymes, overview | Bos taurus |
0.00000013 | - |
mutant M344H in presence of 5 mM Mn2+ | Bos taurus |
0.00000032 | - |
mutant M344E in presence of 5 mM Mn2+ | Bos taurus |
0.00000191 | - |
mutant M344Q in presence of 5 mM Mn2+ | Bos taurus |
0.00000197 | - |
mutant M344S in presence of 5 mM Mn2+ | Bos taurus |
0.00000416 | - |
mutant M344A in presence of 5 mM Mn2+ | Bos taurus |
0.00000535 | - |
wild-type enzyme in presence of 5 mM Mn2+ | Bos taurus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-galactose + N-acetyl-D-glucosamine | - |
Bos taurus | UDP + N-acetyllactosamine | - |
? | |
UDP-galactose + N-acetyl-D-glucosamine | specific for UDP-galactose | Bos taurus | UDP + N-acetyllactosamine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta1,4-galactosyltransferase-I | - |
Bos taurus |
beta4Gal-T1 | - |
Bos taurus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bos taurus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.097 | - |
UDP-galactose | mutant M344H, pH 8.0, 30°C | Bos taurus | |
3.6 | - |
UDP-galactose | wild-type enzyme, pH 8.0, 30°C | Bos taurus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Bos taurus |