Extraction of Membrane Components from Neisseria gonorrhoeae Using Catanionic Surfactant Vesicles: A New Approach for the Study of Bacterial Surface Molecules

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dc.contributor.authorStein, Daniel C.
dc.contributor.authorStocker, Lenea H.
dc.contributor.authorPowell, Abigail E.
dc.contributor.authorKebede, Salsawi
dc.contributor.authorWatts, David
dc.contributor.authorWilliams, Emma
dc.contributor.authorSoto, Nicholas
dc.contributor.authorDhabaria, Avantika
dc.contributor.authorFenselau, Catherine
dc.contributor.authorGanapati, Shweta
dc.contributor.authorDeShong, Philip
dc.date.accessioned2023-11-08T19:20:24Z
dc.date.available2023-11-08T19:20:24Z
dc.date.issued2020-08-20
dc.description.abstractIdentification of antigens is important for vaccine production. We tested extraction protocols using cetyltrimethylammonium tosylate (CTAT) and sodium dodecylbenzenesulfonate (SDBS) to formulate surfactant vesicles (SVs) containing components from Neisseria gonorrhoeae. Carbohydrate and protein assays demonstrated that protein and carbohydrates were incorporated into the vesicle leaflet. Depending on the extraction protocol utilized, 100–400 µg of protein/mL of SVs solution was obtained. Gel electrophoresis followed by silver staining demonstrated that SV extracts contained lipooligosaccharide and a subset of bacterial proteins and lipoproteins. Western blotting and mass spectral analysis indicated that the majority of the proteins were derived from the outer membrane. Mass spectrometric and bioinformatics analysis of SVs identified 29 membrane proteins, including porin and opacity-associated protein. Proteins embedded in the SVs leaflet could be degraded by the addition of trypsin or proteinase K. Our data showed that the incorporation of CTAT and SDBS into vesicles eliminated their toxicity as measured by a THP-1 killing assay. Incorporation of gonococcal cell surface components into SVs reduced toxicity as compared to the whole cell extracts, as measured by cytokine induction, while retaining the immunogenicity. This process constitutes a general method for extracting bacterial surface components and identification of antigens that might be included in vaccines.
dc.description.urihttps://doi.org/10.3390/pharmaceutics12090787
dc.identifierhttps://doi.org/10.13016/dspace/xnkp-lkk2
dc.identifier.citationStein, D.C.; H. Stocker, L.; Powell, A.E.; Kebede, S.; Watts, D.; Williams, E.; Soto, N.; Dhabaria, A.; Fenselau, C.; Ganapati, S.; et al. Extraction of Membrane Components from Neisseria gonorrhoeae Using Catanionic Surfactant Vesicles: A New Approach for the Study of Bacterial Surface Molecules. Pharmaceutics 2020, 12, 787.
dc.identifier.urihttp://hdl.handle.net/1903/31321
dc.language.isoen_US
dc.publisherMDPI
dc.relation.isAvailableAtCell Biology & Molecular Geneticsen_us
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_us
dc.relation.isAvailableAtCollege of Computer, Mathematical & Natural Sciencesen_us
dc.relation.isAvailableAtUniversity of Maryland (College Park, MD)en_us
dc.subjectcatanionic surfactant vesicles
dc.subjectantigen
dc.subjectvaccine
dc.subjectNeisseria gonorrhoeae
dc.subjectmass spectral analysis
dc.titleExtraction of Membrane Components from Neisseria gonorrhoeae Using Catanionic Surfactant Vesicles: A New Approach for the Study of Bacterial Surface Molecules
dc.typeArticle
local.equitableAccessSubmissionNo

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