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Literature summary extracted from

  • Huang, C.; Mohanty, S.; Banerjee, M.
    A novel method of production and biophysical characterization of the catalytic domain of yeast oligosaccharyl transferase (2010), Biochemistry, 49, 1115-1126.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

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

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.4.99.18 endoplasmic reticulum
-
 Saccharomyces cerevisiae 5783
-

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.99.18 Saccharomyces cerevisiae
-
-
-
2.4.99.18 Saccharomyces cerevisiae P39007
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.99.18 yeast oligosaccharyl transferase
-
 Saccharomyces cerevisiae

General Information

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