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Literature summary extracted from

  • Hese, K.; Otto, C.; Routier, F.H.; Lehle, L.
    The yeast oligosaccharyltransferase complex can be replaced by STT3 from Leishmania major (2008), Glycobiology, 19, 160-171.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.99.18 PCR-amplification, FLAG-tag, expression in Saccharomyces cerevisiae Leishmania major
2.4.99.18 STT3 from Leishmania is able to complement stt3 deficiency in yeast during vegetative growth, but only poorly during sporulation. The Leishmania STT3 homolog is functional mainly as a free enzyme with a broad specificity for the glycosyl donor. When incorporated into the OST complex, it accepts the common Glc3Man9GlcNAc2-PP-Dol donor. Three out of the four STT3 paralogs are able to functionally complement not only an stt3 mutant but also ost1, ost2, wbp1, or swp1 mutants. STT3 from Leishmania is not incorporated into the complex and most likely functions as a monomeric enzyme Leishmania major

Protein Variants

EC Number Protein Variants Comment Organism
2.4.99.18 D47E mutant does not support glycosylation in vivo Leishmania major
2.4.99.18 D647A mutant does not support glycosylation in vivo, no growth of yeast cells Leishmania major
2.4.99.18 D706A mutant does not support glycosylation in vivo, no growth of yeast cells Leishmania major
2.4.99.18 D97A mutant does not support glycosylation in vivo, no growth of yeast cells Leishmania major
2.4.99.18 E45E reduced oligosaccharyltransferase activity, sufficient to allow growth of yeast cells Leishmania major
2.4.99.18 E95A mutant does not support glycosylation in vivo, no growth of yeast cells Leishmania major
2.4.99.18 K709A mutant does not support glycosylation in vivo, no growth of yeast cells Leishmania major
2.4.99.18 additional information 3 of the 4 STT3 Leishmania major STT3 proteins (not STT3-3) can complement stt3 deficiency in Saccharomyces cerevisiae during vegetative growth but not during sporulation, addition of osmotic stabilizer sorbitol (1 M) rescues the lethal phenotype during sporulation in rare cases, the yeast enzyme complex subunits ost1, ost2, wbp1 and swp1 can also be complemented by the 3 Leishmania major functional STT3 homologues Leishmania major

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.99.18 92000
-
 recombinant monomeric Leishmania major SST3 enzyme with FLAG-tag, calculated and confirmed by SDS-PAGE Leishmania major
2.4.99.18 180000
-
 non-denaturing PAGE Leishmania major
2.4.99.18 180000
-
 recombinant Leishmania major SST3 with FLAG-tag, BN-PAGE probed with anti-FLAG antiserum Leishmania major
2.4.99.18 480000
-
 oligosaccharyltransferase complex, native polyacrylamide gel electrophoresis Saccharomyces cerevisiae
2.4.99.18 480000
-
 recombinant Leishmania major SST3 (with exception of SST3-3) incorporated in the yeast oligosaccharyltransferase complex when native yeast SST3 is not expressed, BN-PAGE Leishmania major

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 Leishmania major
-
 dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
-
 ?
2.4.99.18 dolichyl diphosphooligosaccharide + [protein]-L-asparagine Saccharomyces cerevisiae
-
 dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.99.18 Leishmania major
-
-
-
2.4.99.18 Leishmania major
-
 This UniProt-ID has been deleted
-
2.4.99.18 Saccharomyces cerevisiae P33767 and Q02791 and P39007 and P41543 and P48439 the enzyme consists of nine subunits. Five of them, Wbp1, Swp1, Stt3, Ost1, and Ost2, are essential for viability of the cell, whereas Ost4, Ost5, Ost3, and Ost6 are not essential but are required for maximal OST activity: Wbp1 (P33767 (SwissProt)), Swp1 (Q02795 (SwissProt)), Stt3 (P39007 (SwissProt)), Ost1 (P41543 (SwissProt)), Ost2 (P46964 (SwissProt)), Ost3 (P48439 (SwissProt)), Ost4 (Q99380 (SwissProt)), Ost5, and Ost6
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.99.18 dolichyl diphosphooligosaccharide + carboxypeptidase Y (CPY) free enzyme Leishmania major dolichyl disphosphate + oligosaccharidyl-(carboxypeptidase Y)
-
 ?
2.4.99.18 dolichyl diphosphooligosaccharide + [protein]-L-asparagine
-
 Leishmania major dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
-
 ?
2.4.99.18 dolichyl diphosphooligosaccharide + [protein]-L-asparagine
-
 Saccharomyces cerevisiae dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
-
 ?
2.4.99.18 dolichyl diphosphooligosaccharide + [protein]-L-asparagine model glycoprotein carboxypeptidase Y. STT3 from Leishmania is able to complement stt3 deficiency in yeast during vegetative growth, but only poorly during sporulation. Transfers efficiently shortened lipid-linked-mannose oligosaccharides. The Leishmania STT3 homolog is functional mainly as a free enzyme with a broad specificity for the glycosyl donor. When incorporated into the OST complex, it accepts the common Glc3Man9GlcNAc2-PP-Dol donor. Three out of the four STT3 paralogs are able to functionally complement not only an stt3 mutant but also ost1, ost2, wbp1, or swp1 mutants Leishmania major dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
-
 ?
2.4.99.18 Glc3Man9GlcNAc2 dolichyl disphosphate + GAYNSTSV microsomes as enzyme source, STT3-1, increase in activity from 51 to 82% when the yeast SST3 expression is turned off, when yeast SST3 is active, only slight change in activity to 103% Leishmania major dolichyl disphosphate + GAY-(Glc3Man9GlcNAc2)NSTSV
-
 ?
2.4.99.18 Man5GlcNAc2 dolichyl diphosphate + GAYNSTSV microsomes as enzyme source, STT3-1, increase in activity from 34 to 109% when the yeast SST3 expression is turned off, when yeast SST3 is active, increase in activity to 115% Leishmania major dolichyl diphosphate + GAY-(Man5GlcNAc2)NSTSV
-
 ?
2.4.99.18 Man6GlcNAc2 dolichyl diphosphate + GAYNSTSV microsomes as enzyme source, STT3-1, increase in activity from 46 to 151% when the yeast SST3 expression is turned off, when yeast SST3 is active, increase in activity to 173% Leishmania major dolichyl diphosphate + GAY-(Man6GlcNAc2)NSTSV
-
 ?
2.4.99.18 Man9GlcNAc2 dolichyl disphosphate + GAYNSTSV microsomes as enzyme source, STT3-1, increase in activity from 54 to 172% when the yeast SST3 expression is turned off, when yeast SST3 is active, increase in activity to 202% Leishmania major dolichyl disphosphate + GAY-(Man9GlcNAc2)NSTSV
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.99.18 monomer 1 * 180000 Leishmania major
2.4.99.18 nonamer the enzyme consists of nine subunits. Five of them, Wbp1, Swp1, Stt3, Ost1, and Ost2, are essential for viability of the cell, whereas Ost4, Ost5, Ost3, and Ost6 are not essential but are required for maximal OST activity: Wbp1 (49392 Da), Swp1 (31653 Da), Stt3 (81529 Da), Ost1 (54072 Da), Ost2 (14698 Da), Ost3, Ost4, Ost5, and Ost6, calculated from sequence Saccharomyces cerevisiae

Synonyms

EC Number Synonyms Comment Organism
2.4.99.18 L.m.STT3
-
 Leishmania major
2.4.99.18 oligosaccharyltransferase
-
 Leishmania major
2.4.99.18 STT3
-
 Leishmania major
2.4.99.18 STT3-1
-
 Leishmania major
2.4.99.18 STT3-2
-
 Leishmania major
2.4.99.18 STT3-3
-
 Leishmania major
2.4.99.18 STT3-4
-
 Leishmania major

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.99.18 24
-
 assay at Leishmania major

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.99.18 7.4
-
 assay at Leishmania major

Expression

EC Number Organism Comment Expression
2.4.99.18 Leishmania major induced by the addition of copper to media up

General Information

EC Number General Information Comment Organism
2.4.99.18 physiological function free monomeric enzyme with broad specificity for nonglucosylated lipid-linked mannose-oligosaccharides typical for protists, incorporated in yeast enzyme complex it also transfers the common eukaryotic Glc3Man9GlcNAc2-PP-Dol donor Leishmania major