ArticleGlycobiology2017
Characterization of the single-subunit oligosaccharyltransferase STT3A from Trypanosoma brucei using synthetic peptides and lipid-linked oligosaccharide analogs.
Article in Glycobiology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed, 34 citations in OpenAlex.
- Structures of ALG3/9/12 reveal the assembly logic of the N-glycan oligomannose core.Nature chemical biology · 2026Article
- GLYCO-BUILD: an enzymatic pipeline for the synthesis of peptides carrying eukaryotic N-glycans.Nature communications · 2025Article
- Recent advances in the biosynthesis of polysaccharide-based antimicrobial glycoconjugate vaccines.Frontiers in microbiology · 2024Review
- Structural and mechanistic studies of the N-glycosylation machinery: from lipid-linked oligosaccharide biosynthesis to glycan transfer.Glycobiology · 2023Review
- Structure, sequon recognition and mechanism of tryptophan C-mannosyltransferase.Nature chemical biology · 2023Article
- Cell-freeFrontiers in molecular biosciences · 2023Article
- Molecular basis for glycan recognition and reaction priming of eukaryotic oligosaccharyltransferase.Nature communications · 2022Article
- Enzymatic Synthesis of Glycans and Glycoconjugates.Advances in biochemical engineering/biotechnology · 2021Review
- Biospektrum : Zeitschrift der Gesellschaft fur Biologishe Chemie (GBCH) und der Vereinigung fur Allgemeine und Angewandte Mikrobiologie (VAAM) · 2021Review
- Transfer learning enables the molecular transformer to predict regio- and stereoselective reactions on carbohydrates.Nature communications · 2020Article
- Synthetic Glycobiology: Parts, Systems, and Applications.ACS synthetic biology · 2020Review
- Article
- Structure of bacterial oligosaccharyltransferase PglB bound to a reactive LLO and an inhibitory peptide.Scientific reports · 2018Article
- Single-pot glycoprotein biosynthesis using a cell-free transcription-translation system enriched with glycosylation machinery.Nature communications · 2018Article
- Biogenesis of Asparagine-Linked Glycoproteins Across Domains of Life-Similarities and Differences.ACS chemical biology · 2018Article
- One pot synthesis of GDP-mannose by a multi-enzyme cascade for enzymatic assembly of lipid-linked oligosaccharides.Biotechnology and bioengineering · 2018Article
- Analysis of substrate specificity ofThe Journal of biological chemistry · 2017Article
- Chemo-enzymatic synthesis of lipid-linked GlcNAc2Man5 oligosaccharides using recombinant Alg1, Alg2 and Alg11 proteins.Glycobiology · 2017Article
- Substrate specificities and reaction kinetics of the yeast oligosaccharyltransferase isoforms.The Journal of biological chemistryArticle
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The initial transfer of a complex glycan in protein N-glycosylation is catalyzed by oligosaccharyltransferase (OST), which is generally a multisubunit membrane protein complex in the endoplasmic reticulum but a single-subunit enzyme (ssOST) in some protists. To investigate the reaction mechanism of ssOST, we recombinantly expressed, purified and characterized the STT3A protein from Trypanosoma brucei (TbSTT3A). We analyzed the in vitro activity of TbSTT3A by synthesizing fluorescently labeled acceptor peptides as well as lipid-linked oligosaccharide (LLO) analogs containing a chitobiose moiety coupled to oligoprenyl carriers of distinct lengths (C10, C15, C20 and C25) and with different double bond stereochemistry. We found that in addition to proline, charged residues at the +1 position of the sequon inhibited glycan transfer. An acidic residue at the -2 position significantly increased catalytic turnover but was not essential, in contrast to the bacterial OST. While all synthetic LLO analogs were processed by TbSTT3A, the length of the polyprenyl tail, but not the stereochemistry of the double bonds, determined their apparent affinity. We also synthesized phosphonate analogs of the LLOs, which were found to be competitive inhibitors of the reaction, although with lower apparent affinity to TbSTT3A than the active pyrophosphate analogs.
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