The structure of an archaeal oligosaccharyltransferase provides insight into the strict exclusion of proline from the N-glycosylation sequon

Taguchi, Yuya; Yamasaki, Tomohiro; Ishikawa, Marie; Kawasaki, Yuki; Yukimura, Ryuji; Mitani, Maki; Hirata, Kunio; Kohda, Daisuke

doi:10.1038/s42003-021-02473-8

Cited by 6 publications

(5 citation statements)

References 62 publications

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“…Sequons in the translated amino acid sequence were identified using a custom-built script coded in Stata 15 [28]code is available in Supplementary File S1. Our code identified sequons with the pattern Asn-Xxx-(Thr/Ser), where the middle position could be any amino acid except proline, since a sequon consisting of Asn-Pro-(Thr/Ser) is not adequate for an N-glycosylation to occur [29]. Sequences were then summarized according to which residues were potentially N-glycosylated.…”

Section: Methodsmentioning

confidence: 99%

Potential Novel N-Glycosylation Patterns Associated with the Emergence of New Genetic Variants of PRRSV-2 in the U.S

et al. 2022

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Glycosylation of proteins is a post-translational process where oligosaccharides are attached to proteins, potentially altering their folding, epitope availability, and immune recognition. In Porcine reproductive and respiratory syndrome virus-type 2 (PRRSV-2), positive selection pressure acts on amino acid sites potentially associated with immune escape through glycan shielding. Here, we describe the patterns of potential N-glycosylation sites over time and across different phylogenetic lineages of PRRSV-2 to better understand how these may contribute to patterns of coexistence and emergence of different lineages. We screened 19,179 PRRSV GP5 sequences (2004–2021) in silico for potential N-glycosylated sites. The emergence of novel combinations of N-glycosylated sites coincided with past PRRSV epidemics in the U.S. For lineage L1A, glycosylation at residues 32, 33, 44, 51, and 57 first appeared in 2012, but represented >62% of all L1A sequences by 2015, coinciding with the emergence of the L1A 1-7-4 strain that increased in prevalence from 8 to 86% of all L1A sequences from 2012 to 2015. The L1C 1-4-4 strain that emerged in 2020 also had a distinct N-glycosylation pattern (residues 32, 33, 44, and 51). From 2020 to 2021, this pattern was responsible for 44–47% of the L1C sequences, contrasting to <5% in years prior. Our findings support the hypothesis that antigenic evolution contributes to the sequential dominance of different PRRSV strains and that N-glycosylation patterns may partially account for antigenic differences amongst strains. Further studies on glycosylation and its effect on PRRSV GP5 folding are needed to further understand how glycosylation patterns shape PRRSV occurrence.

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Section: Methodsmentioning

confidence: 99%

Potential Novel N-Glycosylation Patterns Associated with the Emergence of New Genetic Variants of PRRSV-2 in the U.S

et al. 2022

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“…In the ternary complex, the external loop EL5 of the STT3 subunit is ordered and packs against the luminal domain of STT3, resulting in the formation of a tunnel that connects the binding sites of acceptor and donor substrates 18 , 30 , 43 , 45 , 46 (Fig. 3c ).…”

Section: Resultsmentioning

confidence: 99%

“…In contrast to higher eukaryotes, certain unicellular eukaryotes produce shorter oligosaccharides due to the absence of specific ALG genes in their genomes 14 – 16 . The transfer of the pre-assembled glycan onto secretory proteins is catalyzed by oligosaccharyltransferase (OST), provided these proteins contain an N -glycosylation sequon N-X-S/T 1 – 3 , 17 , 18 . Following glycan transfer, glucosidase-I and glucosidase-II sequentially remove the two terminal glucose units of the newly attached glycan.…”

Section: Introductionmentioning

confidence: 99%

Molecular basis for glycan recognition and reaction priming of eukaryotic oligosaccharyltransferase

et al. 2022

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Oligosaccharyltransferase (OST) is the central enzyme of N-linked protein glycosylation. It catalyzes the transfer of a pre-assembled glycan, GlcNAc2Man9Glc3, from a dolichyl-pyrophosphate donor to acceptor sites in secretory proteins in the lumen of the endoplasmic reticulum. Precise recognition of the fully assembled glycan by OST is essential for the subsequent quality control steps of glycoprotein biosynthesis. However, the molecular basis of the OST-donor glycan interaction is unknown. Here we present cryo-EM structures of S. cerevisiae OST in distinct functional states. Our findings reveal that the terminal glucoses (Glc3) of a chemo-enzymatically generated donor glycan analog bind to a pocket formed by the non-catalytic subunits WBP1 and OST2. We further find that binding either donor or acceptor substrate leads to distinct primed states of OST, where subsequent binding of the other substrate triggers conformational changes required for catalysis. This alternate priming allows OST to efficiently process closely spaced N-glycosylation sites.

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“…Single subunit OST was found in the bacterial pathway of glycoprotein biosynthesis, such as PglB from Campylobacter jejuni (Wacker et al, 2002;Deshpande et al, 2008;Lizak et al, 2011;Jinnelov et al, 2017;Poljak et al, 2017;Mohanty et al, 2020;Schjoldager et al, 2020) and TbSTT3A from Trypanosoma brucei for N-glycan (Wilson et al, 2021). The archaeal pathway also includes the single subunit OST such as AglB (Meyer and Albers, 2014;recent review, see Eichler, 2020), whose X-ray structure from Archaeoglobusfulgidus has been reported very recently (Taguchi et al, 2021). The most studied PglB can transfer the highly conserved heptasaccharide composed of GalNAc 5 Glc 1 Bac 1 (Bac: bacillosamine, 2,4-acetamide-2.4.6-trideoxy-D-glucose) to proteins in the periplasm of C. jejuni.…”

Section: Ost Strategymentioning

confidence: 99%

Recent Chemical and Chemoenzymatic Strategies to Complex-Type N-Glycans

et al. 2022

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Glycosylation is one of the major forms of protein post-translational modification. N-glycans attached to proteins by covalent bonds play an indispensable role in intercellular interaction and immune function. In human bodies, most of the cell surface glycoproteins and secreted glycopeptides are modified with complex-type N-glycans. Thus, for analytical or medicinal purposes, efficient and universal methods to provide homogeneous complex-type N-glycans have been an urgent need. Despite the extremely complicated structures, tremendous progress in the synthesis of N-glycans has been achieved. On one hand, chemical strategies are shown to be effective to prepare core oligosaccharides of N-glycans by focusing on stereoselective glycosylations such as β-mannosylation and α-sialylation, as well as the methodology of the N-glycan assembly. On the other hand, chemoenzymatic strategies have also become increasingly powerful in recent years. This review attempts to highlight the very recent advancements in chemical and chemoenzymatic strategies for eukaryotic complex-type N-glycans.

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The structure of an archaeal oligosaccharyltransferase provides insight into the strict exclusion of proline from the N-glycosylation sequon

Cited by 6 publications

References 62 publications

Potential Novel N-Glycosylation Patterns Associated with the Emergence of New Genetic Variants of PRRSV-2 in the U.S

Potential Novel N-Glycosylation Patterns Associated with the Emergence of New Genetic Variants of PRRSV-2 in the U.S

Molecular basis for glycan recognition and reaction priming of eukaryotic oligosaccharyltransferase

Recent Chemical and Chemoenzymatic Strategies to Complex-Type N-Glycans

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