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  • Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter
    Pszon-Bartosz, Kamila ...
    Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-basedbiomimetic sensor and separation device. In ... this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR) = 50 more than 10(5) FomA proteins could be incorporated in a bilayer array with a total membrane area of 2 mm(2) within 20 min. This novel assay for quantifying protein delivery into lipid bilayers may be a useful tool in developing biomimetic membrane applications.
    Source: Biochemical and biophysical research communications. - ISSN 0006-291X (Vol. 406, no. 1, 2011, str. 96-100)
    Type of material - article, component part
    Publish date - 2011
    Language - english
    COBISS.SI-ID - 75346433
    DOI

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