{
  "abstract": "Lipoprotein and OmpW are two major components of the outer membrane (OM) in Gram-negative bacteria and play essential roles in various physiological processes, e.g., protein secretion, folding, and localization. They typically assist in maintaining client proteins in a folded state or form pore channels during secretion. However, how OM proteins facilitate the secretion of proteins that lack classical signal peptides remains elusive. Here, we demonstrate that they contribute to the secretion of unconventional B-type dye-decolorizing peroxidase (DypB2985) in Pseudomonas putida. The lipoprotein, Lpp1528, which contains a Sec signal peptide, is translocated to the periplasm through the Sec pathway and anchored in the inner leaflet of the OM. Lpp1528 recognizes the C-terminal hydrophobic region of DypB2985 in the cytoplasm and facilitates its coupling to the Sec machinery for inner membrane translocation, despite DypB2985 lacking a canonical N-terminal Sec signal peptide. Following translocation, the two proteins appear to dissociate in the periplasm. Subsequently, another OM protein, OmpW4836, recognizes the N-terminal hydrophobic region of periplasmic DypB2985 and mediates its incorporation into outer membrane vesicles (OMVs) for extracellular delivery. Our study reveals the crosstalk between the Sec pathway and OMVs in the secretion of a non-canonical peroxidase, in which OM proteins, acting as the molecular tethers, mediate the stepwise secretion process. It expands our understanding of non-classical protein secretion mechanisms and bacterial survival strategies. Moreover, the identified OMV sorting mechanism offers potential for the further functionalization of OMVs as versatile biotechnological platforms. Author summary: Bacterial protein secretion is a fundamental physiological process. Proteins commonly contain signal peptides and are exported via classical secretory pathways. However, some proteins without any characterized signal sequences can also leave the cytoplasm and be transported across cell membranes, posing a major unresolved question in the field. Here, we provide insights into the secretion of unconventional B-type dye-decolorizing peroxidases (DypB2985) in Pseudomonas putida. The lipoprotein, Lpp1528, identifies DypB2985 in the cytoplasm and targets it to the Sec secretion components, which transport DypB2985 across the inner membrane. Subsequently, another outer membrane protein, OmpW4836, recognizes periplasmic DypB2985 and mediates its sorting into OMVs for extracellular delivery. Overall, this study advances the understanding of non-classical protein secretion pathways and the inclusion sorting mechanism of OMVs, raising the possibility for the development of tailored biocatalytic and delivery platforms.",
  "abstract_shown": true,
  "affiliations": [
    "Institute of Marine Science and Technology, Shandong University, Qingdao, China",
    "Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Zhuhai, China",
    "Shandong Key Laboratory of Intelligent Marine Engineering Geology, Environment and Equipment, Qingdao, China"
  ],
  "article_type": "Research Article",
  "arxiv": null,
  "author_total": 3,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Congying Liang"
    },
    {
      "affs": [
        1
      ],
      "name": "Wenping Zhu"
    },
    {
      "affs": [
        2
      ],
      "name": "Lu Lin"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/bghgaqaektsvg.bib",
  "cited_by": 0,
  "doi": "10.1371/journal.pgen.1012311",
  "fulltext_url": "https://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1012311",
  "id": "bghgaqaektsvg",
  "issns": [
    "1553-7404"
  ],
  "landing_url": "https://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1012311",
  "language": null,
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1311|Bacterial Genetics and Biotechnology"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Proteins/Lipoproteins/Lipoprotein secretion"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Proteins/Post-translational modification/Signal peptides"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Proteins/Protein transport/Protein secretion"
    ],
    [
      "plos",
      "/Biology and life sciences/Cell biology/Cell processes/Protein transport/Protein secretion"
    ],
    [
      "plos",
      "/Biology and life sciences/Cell biology/Cellular structures and organelles/Cell membranes/Membrane proteins/Outer membrane proteins"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Bacteriology/Bacterial physiology/Secretion systems"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Microbial physiology/Bacterial physiology/Secretion systems"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Periplasm"
    ],
    [
      "plos",
      "/Biology and life sciences/Organisms/Bacteria/Pseudomonas/Pseudomonas putida"
    ],
    [
      "plos",
      "/Biology and life sciences/Physiology/Physiological processes/Secretion"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pathology and laboratory medicine/Pathogens/Virulence factors/Secretion systems"
    ]
  ],
  "openalex": "W7213257671",
  "pdf_url": "https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1012311&type=printable",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-15",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/bghgaqaektsvg.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:49Z",
      "native_id": "10.1371/journal.pgen.1012311",
      "source": "plos",
      "url": "https://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1012311"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "plos:cell biology",
      "subfield": "cell-biology"
    },
    {
      "field": "biology",
      "origin": "plos:microbiology",
      "subfield": "microbiology"
    }
  ],
  "title": "Outer membrane proteins mediate unconventional secretion of Pseudomonas peroxidase through crosstalk between the Sec pathway and outer membrane vesicles",
  "title_text": "Outer membrane proteins mediate unconventional secretion of Pseudomonas peroxidase through crosstalk between the Sec pathway and outer membrane vesicles",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:17:49Z",
  "url": "https://research.draconitic.com/paper/bghgaqaektsvg",
  "venue": "PLOS Genetics",
  "versions": []
}