Abstract
We describe and characterize reconstituted proteolipid vesicles (rPLV) prepared from solubilized Mycoplasma fermentans membranes and studied their binding to and fusion with host Molt-3 cells. The rPLV were prepared following membrane solubilization by Triton X-100 and detergent removal by SM-2 resin beads. The vesicles thus obtained had a rather uniform diameter of about 1 μm and were sealed as monitored by measuring in an assay that measures the quenching by sodium dithionite of a hydrophobic fluorescent probe incorporated into the rPLV membrane. The rPLV adhered to Molt-3 cells and, based on measurements of lipid mixing, fused with the host cells at a similar rate and to about the same extent as intact M. fermentans. Preliminary experiments showed that a chimeric protein, GnRH-PE66, could be encapsulated within these rPLV, opening the way to develop a system for the transfer of high-molecular weight soluble molecules, encapsulated in the rPLV, to target eukaryotic cells.
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Acknowledgments
We thank H. Lorberboum-Galski for providing the chimeric protein GnRH-PE66.
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Rechnitzer, H., Rottem, S. Reconstituted Proteolipid Vesicles Prepared from Mycoplasma fermentans Membranes Are Able to Bind and Fuse with Molt-3 Cells. Curr Microbiol 53, 293–297 (2006). https://doi.org/10.1007/s00284-006-0007-8
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DOI: https://doi.org/10.1007/s00284-006-0007-8