{
  "abstract": "Protein N-glycosylation is catalysed by the oligosaccharyltransferase (OST) complex, yet how local sequon composition controls substrate selection by human OST remains poorly understood. Here we combine atomistic molecular dynamics, alchemical free-energy calculations, in cellulo glycosylation assays and human glycoproteome analysis to define the molecular basis of sequence-dependent N-glycosylation. We built, to our knowledge, the first atomistic model of human OST with both the lipid-linked oligosaccharide donor and an acceptor peptide simultaneously accommodated in a catalytically competent complex. Simulations show that conserved STT3 motifs stabilize the acceptor Asn and explain how substitutions at positions 0, +1 and +2 alter binding and productive geometry. Cellular assays in HEK 293T cells validate the predicted loss of glycosylation for N0Q sequon, +1 Pro and the +2 hierarchy Thr > Ser >> Cys. Analysis of 14,800 human glycosites confirms these rules at proteome scale.",
  "abstract_shown": true,
  "affiliations": [
    "Universitat de Valencia"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "name": "M. Andjelkovic"
    },
    {
      "name": "J. F. Belen-Aguilar"
    },
    {
      "name": "D. Belda"
    },
    {
      "name": "J. J. Ruiz-Pernia"
    },
    {
      "affs": [
        0
      ],
      "name": "I. Tunon"
    },
    {
      "name": "I. Mingarro"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4ja4q7cshmmb6.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.09.03.749169",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.09.03.749169v1.full",
  "id": "4ja4q7cshmmb6",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.09.03.749169v1",
  "language": null,
  "licence": "CC-BY-NC",
  "licence_label": "CC BY-NC",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "biochemistry"
    ],
    [
      "openalex",
      "1312|Glycosylation and Glycoproteins Research"
    ]
  ],
  "openalex": "W7211911822",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.09.03.749169v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-03",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/4ja4q7cshmmb6.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:20:00Z",
      "native_id": "10.64898/2026.09.03.749169",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.09.03.749169v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Glycosylation and Glycoproteins Research",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Molecular Determinants of Sequence-Dependent N-glycosylation by the Human Oligosaccharyltransferase",
  "title_text": "Molecular Determinants of Sequence-Dependent N-glycosylation by the Human Oligosaccharyltransferase",
  "type": "preprint",
  "updated_at": "2026-10-30T12:20:00Z",
  "url": "https://research.draconitic.com/paper/4ja4q7cshmmb6",
  "venue": "bioRxiv",
  "versions": []
}