Abstract
Lipid mixing due to membrane fusion has been measured by monitoring the concentration-dependent properties of membrane-associated probes, usually a fluorescent probe (Struck et al., 1981; Hoekstra et al., 1984). Aside from displaying concentration-dependent fluorescence, probes to be used for fusion assays should 1) not interchange rapidly between membranes, 2) resemble as closely as possible a phospholipid molecule, 3) not interfere with the natural packing and local structure of the bilayer, 4) have fluorescence properties that are insensitive to the presence of added fusogenic agents, and 5) have a high extinction coefficient and quantum yield over broad excitation and emission limits. While the combination of head-group-labeled N-(4-nitro-2,1,3-benzoxadiazol (NBD)- and rhodamine (Rh)-labeled phosphatidylethanolaraines (PE) (Struck, et al., 1983) meets many of these requirements, we (Parente and Lentz, 1986a; 1986b) and others (Morris et al., 1985) have shown that the fluorescence of these probes is quenched by addition of fusogenic agents such as Ca2+ (Figure 1) and poly(ethylene glycol) (PEG). In addition, octadecyl Rhodamine B chloride (R18)> while having certain advantages for the study of biological membrane fusion, has the disadvantages of not resembling a phospholipid and of slowly interchanging between membranes (Hoekstra et al., 1984).
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© 1988 Plenum Press, New York
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Lentz, B.R., Burgess, S.W., Grattont, E. (1988). Concentration Dependence of DPHpPC Fluorescence Lifetime: Photophysics and Utility for Monitoring Membrane Fusion. In: Ohki, S., Doyle, D., Flanagan, T.D., Hui, S.W., Mayhew, E. (eds) Molecular Mechanisms of Membrane Fusion. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-1659-6_41
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DOI: https://doi.org/10.1007/978-1-4613-1659-6_41
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