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Literature summary for 2.4.1.138 extracted from
- Purification of an alpha-N-acetylglucosaminyltransferase from the yeast Kluyveromyces lactis and a study of mutants defective in this enzyme activity (1982), Biochemistry, 21, 1561-1570.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | analysis of mutations in strain mnn2-2 | Kluyveromyces lactis |
General Stability
| General Stability | Organism |
|---|---|
| Triton X-100 stabilizes solubilized enzyme | Kluyveromyces lactis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | strong, 1 mM, even in the presence of Mn2+, wild-type | Kluyveromyces lactis |  |
| Cu2+ | wild-type | Kluyveromyces lactis | |
| imidazole | wild-type | Kluyveromyces lactis | |
| Mn2+ | above 10 mM, wild-type | Kluyveromyces lactis | |
| NaCl | weak | Kluyveromyces lactis | |
| NaN3 | - |
Kluyveromyces lactis | |
| phosphate | wild-type, complete inhibition at 40 mM | Kluyveromyces lactis | |
| Tris | wild-type | Kluyveromyces lactis | |
| Tween 80 | - |
Kluyveromyces lactis | |
| UDP | wild-type | Kluyveromyces lactis | |
| UDP-hexylamine | wild-type | Kluyveromyces lactis | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0364 | - |
UDP-N-acetylglucosamine | mutant 2-2 | Kluyveromyces lactis | |
| 0.0447 | - |
UDP-N-acetylglucosamine | wild-type | Kluyveromyces lactis | |
| 11 | - |
1,3-alpha-D-mannosyl-1,2-alpha-D-mannosyl-1,2-alpha-D-mannosyl-D-mannose | solubilized enzyme | Kluyveromyces lactis | |
| 13 | - |
1,3-alpha-D-mannosyl-1,2-alpha-D-mannosyl-1,2-alpha-D-mannosyl-D-mannose | membrane-bound enzyme | Kluyveromyces lactis | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| endoplasmic reticulum | integral membrane protein | Kluyveromyces lactis | 5783 | - |
| protoplast | - |
Kluyveromyces lactis | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mn2+ | requirement, 10 mM, inhibits at higher concentrations, Mg2+ or Ca2+ cannot replace Mn2+ | Kluyveromyces lactis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 300000 | - |
gel filtration in the presence of Triton X-100, native MW may be lower | Kluyveromyces lactis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UDP-N-acetyl-D-glucosamine + 1,3-alpha-D-mannosyl-1,2-alpha-D-mannosyl-1,2-alpha-D-mannosyl-D-mannose | Kluyveromyces lactis | involved in biosynthesis of mannoprotein side-chain units | UDP + 1,3-alpha-D-mannosyl-1,2-(N-acetyl-alpha-D-glucosaminyl-alpha-D-mannosyl)-1,2-alpha-D-mannosyl-D-mannose | - |
? | |
Organic Solvent Stability
| Organic Solvent | Comment | Organism |
|---|---|---|
| additional information | organic solvent extraction, unstable to | Kluyveromyces lactis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Kluyveromyces lactis | - |
yeast, wild-type strains Y-58a his 4c or his 3 and mutant strains mnn1, mnn2-1 or mnn2-2 | - |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | possesses no N-acetylglucosamine residues, enzyme does not bind to wheat germ agglutinin | Kluyveromyces lactis |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| partial | Kluyveromyces lactis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
activity of wild-type and mutant strains with different acceptors | Kluyveromyces lactis |
| 0.0056 | - |
- |
Kluyveromyces lactis |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, in imidazole-glycerol buffer, 2% Triton X-100, t1/2: 80 days | Kluyveromyces lactis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UDP-N-acetyl-alpha-D-glucosamine + 1,3-alpha-D-mannosyl-1,2-alpha-D-mannosyl-1,2-alpha-D-mannosyl-D-mannose | - |
Kluyveromyces lactis | UDP + 1,3-alpha-D-mannosyl-1,2-(N-acetyl-alpha-D-glucosaminyl-alpha-D-mannosyl)-1,2-alpha-D-mannosyl-D-mannose | - |
? | |
| UDP-N-acetyl-D-glucosamine + 1,3-alpha-D-mannosyl-1,2-alpha-D-mannosyl-1,2-alpha-D-mannosyl-D-mannose | i.e. mannotetraose acceptor: acceptor substrates are mannoprotein, saccharides with more than 25 mannose-units, with 15 to 25 units, with 8 to 15 units, with 5 to 8 units and below 5 units, the latter being the best substrates | Kluyveromyces lactis | UDP + 1,3-alpha-D-mannosyl-1,2-(N-acetyl-alpha-D-glucosaminyl-alpha-D-mannosyl)-1,2-alpha-D-mannosyl-D-mannose | - |
? | |
| UDP-N-acetyl-D-glucosamine + 1,3-alpha-D-mannosyl-1,2-alpha-D-mannosyl-1,2-alpha-D-mannosyl-D-mannose | involved in biosynthesis of mannoprotein side-chain units | Kluyveromyces lactis | UDP + 1,3-alpha-D-mannosyl-1,2-(N-acetyl-alpha-D-glucosaminyl-alpha-D-mannosyl)-1,2-alpha-D-mannosyl-D-mannose | - |
? | |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | - |
2 h, inactivation | Kluyveromyces lactis |
| 45 | - |
t1/2: 4 min | Kluyveromyces lactis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
pI: 4.9, in the presence of Triton X-100 and 2 M urea | Kluyveromyces lactis |
| 6.5 | - |
- |
Kluyveromyces lactis |
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