Abstract
A model is proposed for the study of molecular mechanisms of a low pH-induced interaction of fusion proteins of enveloped viruses and cell membranes. The model consists of large monolamellar liposomes containing ionophore nigericin in their membranes and ectodomains of fusion protein in their inner space. The process of interaction of the protein with the lipid bilayer is triggered by acidification of the liposomal constituents to the pH of fusion with the help of nigericin by adding citric acid to the outer medium. To visualize the protein structural reorganization, the tritium planigraphy was used.
Comparison of the values of specific labelling of the proteins and distribution of radioactivity in individual amino acids in control (at neutral pH) and experimental liposome samples (at the pH of fusion) permits to realise the character of protein-membrane interaction. We have obtained the first results in the study of interaction of the bromelain-released soluble ectodomain of the HAXX molecule (BHA)—with the lipid membrane. The observed increase in the protein specific activity and selective increase in the specific activity of hydrophobic amino acids Ile, Phe and Tyr in experimental liposome samples as compared with the controls did not contradict to the conventional concept, that a hydrophobic N-terminus of HA2 subunit of hemagglutinin is responsible for its interaction with lipid membranes.
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References
Arbatsky, N. P., Lichosherstov, L. M., Medvedev, S. A., Senchencova, S. N., Jurtov, D. V., Derevitskaya, V. A., Kochetkov, N. K. (1983)Doklady AN SSSR,271:1257–1260.
Baratova, L. A., Grebenschchikov, N. I., Shishkov, A. V., Kashirin, I. A., Radavsky, J. L., Jarvekulg, L. V., Saarma, M. J. (1990)J. Gen. Virol. (in press).
Brand, C. M. and Skehel, J. J. (1972)Nature New Biology,238:145–147.
Gedrovitch, A. V., Goldanskii, V. I., Rumyanzev, U. M., Unukovitch, M. M. and Shishkov, A. V. (1984)Radiochimia,26:483–494.
Gedrovitch, A. V., Shishkov, A. V., Goldanskii, V. I., Baratova, L. A., Grebenshchikov, N. I. and Efimov. A. V. (1990)Eur. Biophys. J. (in press).
Gething, M. J., Bye, J., Skehel, J. J. and Waterfield, M. D. (1980)Nature,287:301–306.
Goldanskii, V. I., Kashirin, I. A., Shishkov, A. V., Baratova, L. A. and Grebenshchikov, N. I. (1988)J. Mol. Biol. 201:567–574.
Gordeeva, L. V., Baratova, L. A., Margolis, L. B., Shishkov, A. V. (1989)Biophisika,34:971–975.
Harter, C., James, P., Bachi, T., Semenza, G. and Brunner, J. (1989)J. Biol. Chem.,264:6459–6464.
Hoekstra, D. and Kok, J. W. (1989)Biosci Rep,9:273–305.
Kendall, D. A. and MacDonald, R. C. (1982)J. Biol. Chem.,257:13892–13895.
Kingsbury, D. W. (1966)J. Mol. Biol.,18:195–203.
Kolb, V. A., Kommer, A. and Spirin, A. S. (1990) in: Conf. “Mechanism and Control of Translation”, May 12–17, Noordwijkerhout, The Netherlands.
Laemmli, U. K. (1970)Nature,227:680–685.
Lear, J. D. and DeGrado, W. F. (1987)J. Biol. Chem.,262:6500–6505.
Margolis, L. B., Rosovskaya, I. A. and Skulachev, V. P. (1987)FEBS Lett.,220:288–290.
Marsh, M. (1984)Biochem J.,218:1–10.
Szoka, F. C., Jr. and Papahadjopoulos, F. C. (1978)Proc. Natl. Acad. Sci. USA,75:4194–4198.
White, J., Kielian, M. and Helenius, A. (1983)Q. Rev. Biophys. 16:151–195.
Yusupov, M. M. and Spirin, A. S. (1988) in: “Methods in Enzymology” vol:164:426–439.
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Glushakova, S.E., Ksenofontov, A.L., Fedorova, N.V. et al. A model for the study of the mechanism of a low pH-induced interaction of the virus fusion proteins and cell membranes. Biosci Rep 11, 131–137 (1991). https://doi.org/10.1007/BF01182481
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DOI: https://doi.org/10.1007/BF01182481