An efficient method for reassembly of fusogenic sendai virus envelopes after solubilization of intact virions with triton X‐100

Volsky, David J.; Loyter, Abraham

doi:10.1016/0014-5793(78)80751-0

Cited by 93 publications

(37 citation statements)

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“…This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact. 7247 fusion by Sendai virus (1) and their poor viability after treatment with polyethylene glycol (2).In this study we report the successful coreconstitution of lymphocyte membrane components with Sendai virus envelopes (7,8) (5 Ci/mmol) were from the Radiochemical Center (Amersham, England). Soybean agglutinin was prepared (9) and converted to its 125I-labeled derivative as described (10).…”

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confidence: 99%

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Alteration of lymphocyte surface properties by insertion of foreign functional components of plasma membrane.

Prujansky-Jakobovits¹,

Volsky²,

Loyter³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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The lymphocyte plasma membrane plays a key role in the functions of the immune system. Much effort is being made to understand the correlation between the composition and structure of the lymphocyte plasma membrane and the associated cell functions. Current approaches to this problem are based primarily on studies of isolated membrane constituents and characterization of cell surface structures in intact cells, mainly done on isolated lymphocyte subpopulations.A promising approach involves the transfer of membrane components from one cell type (the donor) to another (the acceptor) in order to study the effect of the new membranal composition on the ability of the acceptor cell to respond to extracellular signals. Present techniques for the transfer of membrane proteins and lipids into plasma membranes of eukaryotic cells involve cell-cell and membrane-cell fusion with fusogenic agents, in particular Sendai virus and polyethylene glycol (1, 2). The transfer of receptors for hormones (3, 4) and asialoglycoproteins (5) to tissue culture cells by these methods has been demonstrated. The methods have also been applied to the transfer of isolated membrane vesicles obtained from highly metastatic cells to the plasma membrane of poorly metastatic sublines (6). However, application of these procedures to the transfer of membrane components to primary cells such as lymphocytes has never been reported, presumably because of the inability of the latter to undergo massive cell-cellThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact. 7247 fusion by Sendai virus (1) and their poor viability after treatment with polyethylene glycol (2).In this study we report the successful coreconstitution of lymphocyte membrane components with Sendai virus envelopes (7,8) (5 Ci/mmol) were from the Radiochemical Center (Amersham, England). Soybean agglutinin was prepared (9) and converted to its 125I-labeled derivative as described (10). Fluorescein isothiocyanate-conjugated limulin was a gift from Michel Monsigny

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mentioning

confidence: 99%

“…In this study we report the successful coreconstitution of lymphocyte membrane components with Sendai virus envelopes (7,8) (5 Ci/mmol) were from the Radiochemical Center (Amersham, England). Soybean agglutinin was prepared (9) and converted to its 125I-labeled derivative as described (10).…”

mentioning

confidence: 99%

Alteration of lymphocyte surface properties by insertion of foreign functional components of plasma membrane.

Prujansky-Jakobovits¹,

Volsky²,

Loyter³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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“…Detergent treatment of Sendal virus particles resulted in solubilization of the two viral envelope glycoproteins, the hemagglutinin/neuraminidase (HN) and the fusion (F) protein [11 ]. These results show that if Sendai virus particles are reduced with DTT before solubllization with Triton X-100, only the F-glycoprotein is extracted into solution.…”

Section: Resultsmentioning

confidence: 95%

“…F-glycoprotein was extracted from Sendai virus particles and membrane vesicles containing the F-glycoprotein were formed as in fig.1 and [11]. RSVE were prepared as in [11].…”

Section: Methodsmentioning

confidence: 99%

Selective extraction of biologically active F‐glycoprotein from dithiothreitol reduced sendai virus particles

Tomasi

Loyter

1981

FEBS Letters

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“…The reassembly of Sendai virus envelopes after solubilization in Triton X-100 was performed essentially as described by Volsky and Loyter [22] with some minor modifications. Prior to detergent-extraction, Sendai virus was washed with 140 mM NaC1, 10 mM Hepes, pH 7.4 (NaCl/Hepes-buffer), by repeated centrifugation at I00000 x g (av) for 30 min at 4°C in the SW 50.1 rotor of a Beckman ultracentrifuge.…”

Section: Dialysis Reconstitution Of Sendai Virus Envelopes With Tritomentioning

confidence: 99%

Reconstitution and fusogenic properties of Sendai virus envelopes

Harmsen¹,

Wilschut²,

Scherphof³

et al. 1985

European Journal of Biochemistry

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Sendai virus membranes were reconstituted by detergent dialysis, using the non-ionic detergents Triton X-100 and octyl glucoside. Membrane reassembly was determined by measuring the surface-density-dependent efficiency of resonance energy transfer between two fluorescent phospholipid analogues, which were co-reconstituted with the viral envelopes. The functional incorporation of the viral proteins was established by monitoring the ability of the reconstitution products to fuse with erythrocyte membranes, utilizing assays based on either resonance energy transfer or on relief of fluorescence selfquenching. The persistent adherence of residual Triton X-100 with the reconstituted membrane was revealed by an artificial detergent-effect on the resonance energy transfer efficiency and the occurrence of hemolysis of human erythrocytes under conditions where fusion does not occur. Properly reconstituted Sendai virus envelopes were obtained with octyl glucoside. The fusion activity of the viral envelopes was dependent on the initial concentration of octyl glucoside used to disrupt the virus and the rate of detergent removal. Rapid removal of detergent by dialysis against large volumes of dialysis buffer (ratio 1 : 850) or by gel filtration produced reconstituted membranes capable of inducing hemagglutination but significant fusion activity was not detected. By decreasing the volume ratio of dialysate versus dialysis buffer to 1 :250 or 1 : 25, fusogenic viral envelopes were obtained. The initial fusion kinetics of the reconstituted viral membrane and the parent virus were different in that both the onset and the initial rate of fusion of the reconstituted membranes were faster, whereas the extents to which both particles eventually fused with the target membrane were similar. The differences in the initial fusion kinetics lead us to suggest that the details of the fusion mechanism between Sendai virus and the target membrane involve factors other than the mere presence of glycoproteins F and HN in the viral bilayer.Finally, the results also indicate that determination of the viral fusion activity in a direct manner, rather than by an indirect assay, such as hemolysis, is imperative for a proper evaluation of the functional properties retained upon viral reconstitution.

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An efficient method for reassembly of fusogenic sendai virus envelopes after solubilization of intact virions with triton X‐100

Cited by 93 publications

References 14 publications

Alteration of lymphocyte surface properties by insertion of foreign functional components of plasma membrane.

Alteration of lymphocyte surface properties by insertion of foreign functional components of plasma membrane.

Selective extraction of biologically active F‐glycoprotein from dithiothreitol reduced sendai virus particles

Reconstitution and fusogenic properties of Sendai virus envelopes

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