2026-10-10T13:51:42Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33816612 2026-09-15T18:18:37Z category_1 category_4 category_8 category_12 category_13 category_14 category_15 category_21 category_24 category_873 category_734 category_61 category_64 category_132 category_134 portal_5 item_type_3 month_year_09_2026

<p>Source data are provided in S1 Data.xlsx.</p> Congying Liang (24951195) Wenping Zhu (11395496) Lu Lin (201289) Biophysics Biochemistry Microbiology Cell Biology Genetics Molecular Biology Neuroscience Biotechnology Evolutionary Biology Chemical Sciences not elsewhere classified Biological Sciences not elsewhere classified Developmental Biology Cancer Infectious Diseases Virology various physiological processes two major components terminal hydrophobic region play essential roles form pore channels div >< p bacterial survival strategies outer membrane vesicles two proteins appear maintaining client proteins sec signal peptide pseudomonas putida </ another om protein inner membrane translocation stepwise secretion process om proteins facilitate outer membrane pseudomonas </ inner leaflet following translocation om proteins sec pathway sec machinery unconventional b typically assist type dye study reveals negative bacteria molecular tethers folded state extracellular delivery <div><p>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 <i>Pseudomonas putida</i>. 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.</p></div> 2026-09-15T18:08:13Z Dataset Dataset 10.1371/journal.pgen.1012311.s004 https://figshare.com/articles/dataset/_p_Source_data_are_provided_in_S1_Data_xlsx_p_/33816612 CC BY 4.0