Cloned (Comment) | Organism |
---|---|
expression of mutants W312C/P401C and W312C/P401C/M340H enzymes in Escherichia coli, the wild-type enzyme forms inclusion bodies after expression in Escherichia coli, overview | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant M340H-Gal-T1 mutant in the open conformation as apo-enzyme, and its Mn2+ and Mn2+-UDP-Gal-bound complexes, hanging-drop vapor diffusion method using a protein solution at 10 mg/ml concentration with a precipitant containing sodium citratebuffer, pH 5.5, and 10% w/v PEG-4000, the crystals of the Mn2+ complex are grown in the droplet containing 10 mM MnCl2, while the crystals containing UDP-Gal and Mn2+ are grown in the presence of 17 mM each of MnCl2 and UDPยGal, X-ray diffraction structure determination and analysis at 1.7-2.0 A resolution | Homo sapiens |
purified recombinant pentenary complex of bovine M344H-Gal-T1 mutant-Mn2+-UDP-GalNAc-Glc-alpha-lactalbumin, hanging-drop vapor diffusion method, using 20 mg/ml of beta-M344H-Gal-T1 and 10 mg/ml of mouse alpha-lactalbumin in the presence of 10 mM each ofUDP-Gal and CaCl2, with the precipitant containing 100 mM NaCl, 100 mM Mes-NaOH buffer, pH 6.0, and 12.5% PEG 4000, the crystals of the complex cannot be obtained in the absence of CaCl2, X-ray diffraction structure determination and analysis at 1.9-2.0 A resolution | Bos taurus |
Protein Variants | Comment | Organism |
---|---|---|
M344H | site-directed mutagenesis, substrate binding structure in comparison to the wild-type enzyme, overview | Bos taurus |
W312C/P401C | site-directed mutagenesis, the mutations facilitate invitro folding of the recombinantly expressed enzyme | Homo sapiens |
W312C/P401C/M340H | site-directed mutagenesis, substrate binding structure in comparison to the wild-type enzyme, overview | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Bos taurus | |
Mn2+ | required, binds first in the catalytic cycle | Bos taurus | |
Mn2+ | required, binds first in the catalytic cycle, Met340 is involved | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-galactose + N-acetyl-beta-D-glucosaminylglycopeptide | Homo sapiens | - |
UDP + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminylglycopeptide | - |
? | |
UDP-galactose + N-acetyl-beta-D-glucosaminylglycopeptide | Bos taurus | - |
UDP + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminylglycopeptide | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bos taurus | - |
- |
- |
Homo sapiens | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
UDP-alpha-D-galactose + N-acetyl-beta-D-glucosaminylglycopeptide = UDP + beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosaminylglycopeptide | reaction mechanism and structural changes | Homo sapiens | |
UDP-alpha-D-galactose + N-acetyl-beta-D-glucosaminylglycopeptide = UDP + beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosaminylglycopeptide | reaction mechanism and structural changes | Bos taurus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-galactose + N-acetyl-beta-D-glucosaminylglycopeptide | - |
Homo sapiens | UDP + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminylglycopeptide | - |
? | |
UDP-galactose + N-acetyl-beta-D-glucosaminylglycopeptide | - |
Bos taurus | UDP + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminylglycopeptide | - |
? | |
UDP-galactose + N-acetyl-beta-D-glucosaminylglycopeptide | catalytic cycle: Mn2+ binds first before UDP-Gal, two flexible loops, a long and a short loop, change their conformation from open to closed | Homo sapiens | UDP + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminylglycopeptide | - |
? | |
UDP-galactose + N-acetyl-beta-D-glucosaminylglycopeptide | catalytic cycle: Mn2+ binds first before UDP-Gal, two flexible loops, a long and a short loop, change their conformation from open to closed | Bos taurus | UDP + beta-D-galactosyl-1,4-N-acetyl-beta-D-glucosaminylglycopeptide | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure analysis, in vivo and invitro folding analysis, open and closed conformation change of the enzyme upon Mn2+ and substrate binding, overview | Homo sapiens |
More | structure analysis, open and closed conformation change of the enzyme upon Mn2+ and substrate binding, overview | Bos taurus |
Synonyms | Comment | Organism |
---|---|---|
beta-1,4-galactosyltransferase-I | - |
Homo sapiens |
beta-1,4-galactosyltransferase-I | - |
Bos taurus |
Gal-T1 | - |
Homo sapiens |
Gal-T1 | - |
Bos taurus |