EC Number | Cloned (Comment) | Organism |
---|---|---|
2.4.99.18 | PCR-amplification, expression of the pure C-terminal domain of the subunit Stt3p in Escherichia coli BL21 (DE3) | Saccharomyces cerevisiae |
2.4.99.18 | subcloning, overexpression, and a method of production of the pure C-terminal domain of Stt3p at 60-70 mg/l in Escherichia coli | Saccharomyces cerevisiae |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.4.99.18 | D518E | the replacement of a key residue, Asp518, located within the WWDYG signature motif (residues 516-520), leads to a distinct tertiary structure, even though both proteins have similar overall secondary structures, as demonstrated by CD, fluorescence and NMR spectroscopies. Asp518 plays a critical structural role, in addition to the catalytic role. The activity of the protein is confirmed by saturation transfer difference and nuclear magnetic resonance titration studies | Saccharomyces cerevisiae |
2.4.99.18 | D518E | within WWDYG signature motif leads to distinct tertiary structure interfering with function (complete loss of N-linked glycosylation activity), lethal | Saccharomyces cerevisiae |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.4.99.18 | additional information | - |
additional information | dissociation constant KD=9.97 mM | Saccharomyces cerevisiae |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
2.4.99.18 | endoplasmic reticulum | - |
Saccharomyces cerevisiae | 5783 | - |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.4.99.18 | 31500 | - |
MALDI-TOF result in accordance with calculated results for histidine-tagged protein subunit Stt3p | Saccharomyces cerevisiae |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.4.99.18 | dolichyl diphosphooligosaccharide + protein L-asparagine | Saccharomyces cerevisiae | - |
dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine | - |
? | |
2.4.99.18 | dolichyl diphosphooligosaccharide + [protein]-L-asparagine | Saccharomyces cerevisiae | oligosaccharyl transferase (OT) is a multisubunit enzyme that catalyzes N-linked glycosylation of nascent polypeptides in the lumen of the endoplasmic reticulum | dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.4.99.18 | Saccharomyces cerevisiae | - |
- |
- |
2.4.99.18 | Saccharomyces cerevisiae | P39007 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.4.99.18 | C-terminal domain of Stt3p, wild-type and mutant D518E | Saccharomyces cerevisiae |
2.4.99.18 | dissolution in denaturing buffer (6 M guanidine hydrochloride, 500 mM Na Cl, 25 mM imidazole, 20 mM phosphate buffer, pH 7.4), cenrtrifugation, supernatant loaded onto nickel-NTA column for affinity chromatography with 20 mM phosphate buffer, pH 6.5, SDS-PAGE | Saccharomyces cerevisiae |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.4.99.18 | dolichyl diphosphooligosaccharide + asparagine-asparagine-threonine-NH2 acceptor peptide | - |
Saccharomyces cerevisiae | dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine | - |
? | |
2.4.99.18 | dolichyl diphosphooligosaccharide + protein L-asparagine | - |
Saccharomyces cerevisiae | dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine | - |
? | |
2.4.99.18 | dolichyl diphosphooligosaccharide + [protein]-L-asparagine | oligosaccharyl transferase (OT) is a multisubunit enzyme that catalyzes N-linked glycosylation of nascent polypeptides in the lumen of the endoplasmic reticulum | Saccharomyces cerevisiae | dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.4.99.18 | yeast oligosaccharyl transferase | - |
Saccharomyces cerevisiae |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.4.99.18 | malfunction | congenital disorders of glycosylation (CDG) have severe effects in humans, complete loss is lethal for eukaryots | Saccharomyces cerevisiae |
2.4.99.18 | physiological function | N-linked glycosylation of nascent polypeptides in the lumen of the endoplasmic reticulum | Saccharomyces cerevisiae |