Reconstitution of fusogenic Sendai virus envelopes by the use of the detergent chaps.
Sendai virus envelopes have been a useful tool in studying the mechanism of membrane-membrane fusion and have served as a vehicle for introducing foreign molecules (e.g., membrane proteins) into recipient cells. Reconstituted Sendai virus envelopes are routinely obtained following solubilization of virus particles with Triton X-100. This detergent has a low critical micellar concentration which precludes it from being the best detergent of choice in reconstitution studies. Nevertheless, it has remained in use since other detergents such as sodium deoxycholate and sodium cholate rendered the resultant vesicles inactive. Triton X-100 may be suboptimal for studies of some proteins that need be coreconstituted with the viral envelopes. Thus, alternative advantageous detergents, which retain the envelope fusogenic activity, have been sought. In this study we show that the synthetic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (Chaps) effectively solubilizes the Sendai virions, and that the vesicles formed by simple reconstitution protocols appear structurally and biochemically similar to those obtained with Triton X-100. The resultant vesicles retain functional integrity as assessed in both fusion and hemolysis assays. This protocol seems to be useful in sendai envelope-mediated reimplantation of Fc epsilon receptors into the plasma membranes of rat basophilic leukemia cells.
D002462:Cell Membrane / Q000032:analysis; D002793:Cholic Acids / Q000494:pharmacology*; D003902:Detergents / Q000494:pharmacology*; D006461:Hemolysis; D017830:Octoxynol; D010222:Parainfluenza Virus 1, Human / Q000032:analysis*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012995:Solubility; D013194:Staining and Labeling; D013501:Surface-Active Agents / Q000494:pharmacology*; D014759:Viral Envelope Proteins / Q000032:analysis*; D014760:Viral Fusion Proteins / Q000032:analysis*
D002793:Cholic Acids; D003902:Detergents; D013501:Surface-Active Agents; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate