| Title | Profiling Protein Citrullination in Extracellular Vesicles by Single-Molecule Detection Using Direct Stochastic Optical Reconstruction Microscopy |
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| Type | Journal article |
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| Authors | Needham, S.R., Davis, B.M., Uysal Onganer, P., Rolfe, D.J., Hristova, M., Kraev, I., Inal, J.M. and Lange, S. |
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| Abstract | Extracellular vesicles (EVs) are critical in cellular communication and pathological biomarkers. Post-translationally deiminated/citrullinated proteins are reported in EV cargoes by LC–MS/MS but it is unknown in which EV sub-types they are exported, as part of EVs' intraluminal cargo or on the EV surface. Here, dSTORM super-resolution microscopy is used to co-localise total citrullinated proteins (pan-Cit), and citrullinated histone H3 (CitH3) to EV subtypes of three cancer cell lines, captured by tetraspanin trio (TT) or phosphatidylserine (PS). Permeabilised and non-permeabilised EVs are analysed with a Bayesian framework using beta-distributed posteriors for binomial outcomes. Pan-Cit and CitH3 labelling is confirmed in EVs as intraluminal cargo and on the EV surface, with higher levels detected in the permeabilized EVs. Pan-Cit staining is higher in TT-bound EVs, but CitH3 staining higher in PS-bound EVs. This study expands the landscape of EV-associated post-translational modifications with translational potential for EV-citrullinome based liquid biopsy tools. |
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| Keywords | Bayesian models |
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| citrullinated histone H3 |
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| citrullination |
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| deimination |
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| dSTORM |
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| extracellular vesicles |
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| liquid biopsy |
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| single-molecule detection |
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| Article number | e202500483 |
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| Journal | Journal of Biophotonics |
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| Journal citation | 2025 |
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| ISSN | 1864-0648 |
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| Year | 2025 |
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| Publisher | Wiley |
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| Publisher's version | License CC BY 4.0 File Access Level Open (open metadata and files) |
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| Digital Object Identifier (DOI) | https://doi.org/10.1002/jbio.202500483 |
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| Web address (URL) | https://onlinelibrary.wiley.com/doi/10.1002/jbio.202500483 |
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| Publication dates |
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| Published | 27 Nov 2025 |
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| Supplemental file | File Access Level Open (open metadata and files) |
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| Funder | UKRI (UK Research and Innovation) |
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