Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Saccharomyces cerevisiae | Oryctolagus cuniculus |
Crystallization (Comment) | Organism |
---|---|
the DELTA270 enzyme crystallizes at 7.5 mg/ml from solutions containing 1 mM UDP-glucose, 1 mM MnCl2, 100 mM sodium acetate, pH 4.54.7, and 1013% (w/v) PEG 4000 in the space group P64, with cell dimensions a = b = 75.0, c = 233.4, gamma = 120° and a dimer in the asymmetric unit. All crystals are grown using hanging drop vapor diffusion methods at 23°C | Oryctolagus cuniculus |
Protein Variants | Comment | Organism |
---|---|---|
D159N | exists as both tetrameric and dimeric species, compared to wild-type enzyme which exists to more than 95% as dimer, self-glucosylation activities below the limit of detection of the assay. Ability to catalyze the transglucosylation of maltose is reduced by 260fold, hydrolysis of UDP-glucose is reduced 12fold | Oryctolagus cuniculus |
D159S | stable enzyme, self-glucosylation activities below the limit of detection of the assay. Transglucosylation activity of the mutant enzyme is reduced to undetectable levels, activity for the hydrolysis of UDP-glucose is reduced 14fold | Oryctolagus cuniculus |
D162N | exists as both tetrameric and dimeric species, compared to wild-type enzyme which exists to more than 95% as dimer, self-glucosylation activities below the limit of detection of the assay, undetectable activity for the transglucosylation of maltose and the hydrolysis of UDP-glucose to free glucose | Oryctolagus cuniculus |
D162S | stable enzyme, self-glucosylation activities below the limit of detection of the assay. 30fold less active for the trans-glucosylation of maltose and 340fold less active for the hydrolysis of UDP-glucose | Oryctolagus cuniculus |
DELTA270-332 | mutant enzyme is fully active, specific activity for self- or transglucosylation is indistinguishable from the full-length enzyme | Oryctolagus cuniculus |
DELTA270-332/D159S | inactive mutant enzyme | Oryctolagus cuniculus |
DELTA270-332/D162N | exists as both tetrameric and dimeric species, compared to wild-type enzyme which exists to more than 95% as dimer | Oryctolagus cuniculus |
DELTA270-332/D162S | 18fold less active for the transglucosylation of maltose and 190fold less active for the hydrolysis of UDP-glucose than wild-type enzyme, activity for the hydrolysis of UDP-glucose is reduced 4fold | Oryctolagus cuniculus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
58000 | - |
truncation mutant enzymes DELTA270-332/D159S and DELTA270-332/D162S, gel filtration | Oryctolagus cuniculus |
103000 | - |
wild-type enzyme in glycosylated and nonglycosylated form, full-length mutant enzymes D162S and D159S, gel filtration | Oryctolagus cuniculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Oryctolagus cuniculus | P13280 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | - |
Oryctolagus cuniculus |
Purification (Comment) | Organism |
---|---|
wild-type and mutant enzymes | Oryctolagus cuniculus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
muscle | - |
Oryctolagus cuniculus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-glucose + glycogenin | self-glucosylation | Oryctolagus cuniculus | UDP + glucosylglycogenin | - |
? | |
UDP-glucose + maltose | trans-glucosylation. 93% of the transferred glucose molecules appears in maltotriose, 6% are attached to glycogenin, and 1% is liberated as free glucose | Oryctolagus cuniculus | UDP + maltotriose | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | truncation mutant enzymes DELTA270-332/D159S and DELTA270/D162S, wild-type enzyme in glycosylated and nonglycosylated form, full-length mutant enzymes D162S and D159S exists as more than 95% dimer. Mutant enzymes D159N, D162N and DELTA270-332/D162N exist as both tetrameric and dimeric species | Oryctolagus cuniculus |
tetramer | mutant enzymes D159N, D162N and DELTA270-332/D162N exist as both tetrameric and dimeric species | Oryctolagus cuniculus |
Synonyms | Comment | Organism |
---|---|---|
glycogenin | - |
Oryctolagus cuniculus |