Kinetics of Sendai virus envelope fusion with erythrocyte membranes and virus-induced hemolysis

Lyles, Douglas S.; Landsberger, Frank R.

doi:10.1021/bi00590a011

Cited by 54 publications

(24 citation statements)

References 39 publications

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“…However, in addition to this statistical rationale, it is necessary to consider theoretical kinetic analysis. These analyses indicate that, although empirical modelling of the data could allow fitting to different equations, in the case of virus-cell fusion (Nir et al, 1986a,b) there is one theoretical singleexponential model that will best explain the fusion process (Lyles and Landsberger, 1979;Tsao and Huang, 1986;Nir et al, 1986a,b;Pedroso de Lima et al, 1991, 1992Ramalho-Santos et al, 1993). All of the dequenching curves obtained in the course of our investigations gave the best fit to the proposed model of the sum of two exponentials, regardless of the assay conditions (pH, temperature, ghost concentration, etc.…”

Section: Discussionmentioning

confidence: 99%

Fusion between Newcastle disease virus and erythrocyte ghosts using octadecyl Rhodamine B fluorescence assay produces dequenching curves that fit the sum of two exponentials

Cobaleda

García‐Sastre

Villar

1994

Biochemical Journal

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The kinetics of fusion between Newcastle disease virus and erythrocyte ghosts has been investigated with the octadecyl Rhodamine B chloride assay [Hoekstra, De Boer, Klappe, and Wilschut (1984) Biochemistry 23, 5675-5681], and the data from the dequenching curves were fitted by non-linear regression to currently used kinetic models. We used direct computer-assisted fitting of the dequenching curves to the mathematical equations. Discrimination between models was performed by statistical analysis of different fits. The experimental data fit the exponential model previously published [Nir, Klappe, and Hoekstra (1986) Biochemistry 25, 2155-2161] but we describe for the first time that the best fit was achieved for the sum of two exponential terms: A1[1-exp(-k1t)]+A2[1-exp(-k2t)]. The first exponential term represents a fast reaction and the second a slow dequenching reaction. These findings reveal the existence of two independent, but simultaneous, processes during the fusion assay. In order to challenge the model and to understand the meaning of both equation, fusion experiments were carried out under different conditions well known to affect viral fusion (changes in pH, temperature and ghost concentration, and the presence of disulphide-reducing agents or inhibitors of viral neuraminidase activity), and the same computer fitting scheme was followed. The first exponential equation represents the viral protein-dependent fusion process itself, because it is affected by the assay conditions. The second exponential equation accounts for a nonspecific reaction, because it is completely independent of the assay conditions and hence of the viral proteins. An interpretation of this second process is discussed in terms of probe transfer between vesicles.

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Section: Discussionmentioning

confidence: 99%

Fusion between Newcastle disease virus and erythrocyte ghosts using octadecyl Rhodamine B fluorescence assay produces dequenching curves that fit the sum of two exponentials

Cobaleda

García‐Sastre

Villar

1994

Biochemical Journal

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“…It is possible that other factors contribute as well to a limited fusion capacity of viruses per ghost. Alterations of membrane fluidity owing to virus-induced cross-linking of cell membrane receptors (1,19,21) and the rate of viral protein diffusion (2, 18) from the fusion areas may additionally affect the fusion ability of viral particles which are about to fuse, particularly during the early stages of the fusion process. However, both these possibilities do not mutually exclude the existence of specific fusion sites on the ghost surface.…”

Section: Discussionmentioning

confidence: 99%

Sendai virus-erythrocyte membrane interaction: quantitative and kinetic analysis of viral binding, dissociation, and fusion

Hoekstra¹,

Klappe²

1986

J Virol

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A kinetic and quantitative analysis of the binding and fusion of Sendai virus with erythrocyte membranes was performed by using a membrane fusion assay based on the relief of fluorescence self-quenching. At 37°C, the process of virus association displayed a half time of 2.5 min; at 4°C, the half time was 3.0 min. The fraction of the viral dose which became cell associated was independent of the incubation temperature and increased with increasing target membrane concentration. On the average, one erythrocyte ghost can accommodate ca. 1,200 Sendai virus particles. The stability of viral attachment was sensitive to a shift in temperature: a fraction of the virions (ca. 30%), attached at 40C, rapidly (half time, ca. 2.5 min) eluted from the cell surface at 37°C, irrespective of the presence of free virus in the medium. The elution can be attributed to a spontaneous, temperature-induced release, rather than to viral neuraminidase activity. Competition experiments with nonlabeled virus revealed that viruses destined to fuse do not exchange with free particles in the medium but rather bind in a rapid and irreversible manner. The fusion rate of Sendai virus was affected by the density of the virus particles on the cell surface and became restrained when more than 170 virus particles were attached per ghost. In principle, all virus particles added displayed fusion activity. However, at high virus-to-ghost ratios, only a fraction actually fused, indicating that a limited number of fusion sites exist on the erythrocyte membrane. We estimate that ca. 180 virus particles maximally can fuse with one erythrocyte ghost.

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“…Influenza virus, on the other hand, did not transfer a membrane-inserted phosphatidylcholine spin label to interacting erythrocytes. These studies provide a dramatic demonstration of transfer of paramyxovirus membrane lipids to the membrane of the erythrocyte with which the virus fuses, an observation that was confirmed in similar studies by Lyles and Landsberger (1979). An electron microscopic confirmation of paramyxovirus membrane fusion with multilayered liposomes was also provided by Haywood (1974).…”

Section: 11a Paramyxovirusesmentioning

confidence: 52%

Cytopathic Effects of Viruses: A General Survey

Wagner

1984

Viral Cytopathology

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Kinetics of Sendai virus envelope fusion with erythrocyte membranes and virus-induced hemolysis

Cited by 54 publications

References 39 publications

Fusion between Newcastle disease virus and erythrocyte ghosts using octadecyl Rhodamine B fluorescence assay produces dequenching curves that fit the sum of two exponentials

Fusion between Newcastle disease virus and erythrocyte ghosts using octadecyl Rhodamine B fluorescence assay produces dequenching curves that fit the sum of two exponentials

Sendai virus-erythrocyte membrane interaction: quantitative and kinetic analysis of viral binding, dissociation, and fusion

Cytopathic Effects of Viruses: A General Survey

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