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J Wilschut

Publications and source records attributed to J Wilschut.

95 records · Page 6Linked to original sources
PubMed1980

Mass action kinetics of phosphatidylserine vesicle fusion as monitored by coalescence of internal vesicle volumes.

The kinetics of Ca2+-induced fusion of sonicated phosphatidylserine vesicles is analyzed by means of the mass action model. The results of calculations are shown to simulate the experimental results for the mixing of aqueous vesicle volumes, release of vesicle contents and for the observed increase in light scattering [Wilschut, J., Düzgünes, N., Fraley, R., & Papahadjopoulos, D. (1980) Biochemistry (first of three papers in this issue)]. The calculations give the distribution of vesicle sizes during the initial stages of the fusion process and an estimate for the occurrence of multiple fusion events. It is estimated that during the first few seconds from the beginning of the fusion process in the above systems only a small fraction of the material trapped will leak during each fusion event. The fraction of material which leaks per fusion event is further reduced with increased Ca2+ concentrations. The values of the rates of fusion which describe the above experiments suggest that the rate limiting step of the overall fusion reaction is the aggregation and close approach of vesicles to each other rather than the fusion event per se.

Calcium↗
PubMed1979

Ca2+-induced fusion of phospholipid vesicles monitored by mixing of aqueous contents.

Ca2+ has a central role in various cellular phenomena involving membrane fusion. However, little is known about the mechanisms involved. Model membrane systems such as phospholipid vesicles have been used extensively to study the mechanism of membrane fusion at the molecular level. For example, phosphatidylserine (PS) vesicles have been shown to undergo massive aggregation and structural rearrangements on additon of Ca2+, with eventual formation of large cochleate structures. Although these structures do not retain appreciable internal volume, their formation has been proposed to result from fusion of the initial vesicles. The significance of the PS--Ca2+ system as a model for biological membrane fusion has been questioned recently by Ginsberg. Based on the observation that divalent cations induce the release of contents from PS vesicles but fail to bring about the uptake of a marker from the medium, he proposes that the vesicles are ruptured completely during interaction with divalent cations and reassemble subsequently to form large non-vesicular structures. The present study demonstrates that the question raised by Ginsberg is not particularly relevant to the phenomenon concerned, and that his experimental observations do not allow the exclusive conclusion that Ca2+ induces lysis of PS vesicles rather than fusion.

Calcium↗
PubMedDate not supplied

Membrane fusion: lipid vesicles as a model system.

In many cellular functions the process of membrane fusion is of vital importance. It occurs in a highly specific and strictly controlled fashion. Proteins are likely to play a key role in the induction and modulation of membrane fusion reactions. Aimed at providing insight into the molecular mechanisms of membrane fusion, numerous studies have been carried out on model membrane systems. For example, the divalent-cation induced aggregation and fusion of vesicles consisting of negatively charged phospholipids, such as phosphatidylserine (PS) or cardiolipin (CL), have been characterized in detail. It is important to note that these systems largely lack specificity and control. Therefore conclusions derived from their investigation can not be extrapolated directly to a seemingly comparable counterpart in biology. Yet, the study of model membrane systems does reveal the general requirements of lipid bilayer fusion. The most prominent barrier to molecular contact between two apposing bilayers appears to be due to the hydration of the polar groups of the lipid molecules. Thus, dehydration of the bilayer surface and fluctuations in lipid packing, allowing direct hydrophobic interactions, are critical to the induction of membrane fusion. These membrane alterations are likely to occur only locally, at the site of intermembrane contact. Current views on the way membrane proteins may induce fusion under physiological conditions also emphasize the notion of local surface dehydration and perturbation of lipid packing, possibly through penetration of apolar amino acid segments into the hydrophobic membrane interior.

Cell Fusion↗
PubMedDate not supplied

Fusion of Semliki Forest virus with cholesterol-containing liposomes at low pH: a specific requirement for sphingolipids.

Semliki Forest virus (SFV) utilizes a membrane fusion strategy to introduce its genome into the host cell. After binding to cell-surface receptors, virus particles are internalized through receptor-mediated endocytosis and directed to the endosomal cell compartment. Subsequently, triggered by the acid pH in the lumen of the endosomes, the viral envelope fuses with the endosomal membrane. As a result of this fusion reaction the viral RNA gains access to the cell cytosol. Low-pH-induced fusion of SFV, in model systems as well as in cells, has been demonstrated previously to be strictly dependent on the presence of cholesterol in the target membrane. In this paper, we show that fusion of SFV with cholesterol-containing liposomes depends on sphingomyelin (SM) or other sphingolipids in the target membrane, ceramide representing the sphingolipid minimally required for mediating the process. The action of the sphingolipid is confined to the actual fusion event, cholesterol being necessary and sufficient for low-pH-dependent binding of the virus to target membranes. The 3-hydroxyl group on the sphingosine backbone plays a key role in the SFV fusion reaction, since 3-deoxy-sphingomyelin does not support the process. This, and the remarkably low levels of sphingolipid required for half-maximal fusion (1-2 mol%), suggest that the sphingolipid does not play a structural role in SFV fusion, but rather acts as a cofactor, possibly through activation of the viral fusion protein. Domain formation between cholesterol and sphingolipid, although it may facilitate SFV fusion, is unlikely to play a crucial role in the process.

Animals↗