The Reconstitution of Biologically Active Glycoproteins into Large Liposomes: Use as a Delivery Vehicle to Animal Cells

  • Susan Gould-Fogerite
  • Joseph E. Mazurkiewicz
  • Donna Bhisitkul
  • Raphael J. Mannino


Two related methods for the efficient reconstitution of membrane proteins into large (0.1 to 4 micron diameter) phospholipid vesicles with a large, internal aqueous space have been developed. These procedures utilize the nonionic detergent ß-D-octyl-glucopyranoside, which is rapidly and easily removed, and do not involve exposure to organic solvents, sonication, or extremes of pH, temperature, or pressure. The methods have been applied to the glycoproteins of influenza (A/PR8/34) and Sendai (parainfluenza type I) viruses. The tight association of the proteins with the lipid bilayer was indicated by their migration with the phospholipid to the top of metrizamide density gradients after sonication, whereas unencapsulated FITC-Dextran remained at the bottom of the gradient. Negative staining and electron microscopy showed peplomers on the surface of the vesicles which were indistinguishable from those seen on the respective viruses. The reconstituted glycoproteins have been shown to retain their biological activities of receptor binding and membrane fusion as assayed by a variety of criteria which include the ability to agglutinate and lyse red blood cells. In addition, binding and delivery of vesicle contents to the cytoplasm of virtually every cell in culture has been demonstrated using fluorescently labeled dextran and immunoglobulin. Consistent with their biological activities in the viruses, vesicles prepared with Sendai virus glycoproteins deliver their contents gradually, without manipulation. In contrast, rapid large scale delivery by vesicles prepared with Influenza virus glycoproteins is activated by brief exposure of cell-bound vesicles to low pH.


Hemolytic Activity Large Vesicle Nucleocapsid Protein Direct Addition Phospholipid Vesicle 
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Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Susan Gould-Fogerite
    • 1
  • Joseph E. Mazurkiewicz
    • 2
  • Donna Bhisitkul
    • 1
  • Raphael J. Mannino
    • 1
  1. 1.Department of Microbiology and ImmunologyAlbany Medical College of Union UniversityAlbanyUSA
  2. 2.Department of AnatomyAlbany Medical College of Union UniversityAlbanyUSA

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