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Separation of measles virus components by equilibrium centrifugation in CsCl gradients

II. Studies on the large and the small hemagglutinin

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Summary

CsCl equilibrium density centrifugation studies have been undertaken with the rapid-sedimenting large hemagglutinin (HA) and slow-sedimenting — “native” — small HA fractions of measles HA. The former appeared rather homogeneous, biological activities being concentrated to fractions with densities of 1.20 to 1.25 g/cc. Nucleic acid determinations by tritium labelling and colorimetric reactions revealed an accumulation of RNA in the same range of densities, whereas no significant amounts of DNA could be detected. A considerable fraction of the RNA present in preparations of the large HA could be eliminated by adsorption with monkey erythrocytes. However, if the material was first treated with Tween 80 and ether no change in RNA content was demonstrable after erythrocyte adsorption.

The “native” small HA appeared inhomogeneous in the gradients. Hemagglutinating activity was spread over a range of densities from 1.25 to 1.12 g/cc. and also occurred as a narrow band at a density of 1.30 g/cc. The location of the latter band corresponds to that of the only peak of activities appearing in Tween and ether treated material, that is an artificially produced small HA. The small amounts of HL and CF activity present in “native” small HA appeared at densities lower than 1.20 g/cc. Although being non-infectious these fractions exerted a rapidly appearing cytopathic effect in tissue cultures. No intracellular viral antigen could be detected by the indirect immuno-fluorescent technique in syncytia formed in this way.

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Authors and Affiliations

  1. Department of Virus Research, Karolinska Institutet, School of Medicine, Stockholm, Sweden

    E. C. J. Norrby, Pernilla Magnusson, L. -G. Falksveden & Margareta Grönberg

  2. Department of Chemistry, National Bacteriological Laboratory, Stockholm, Sweden

    E. C. J. Norrby, Pernilla Magnusson, L. -G. Falksveden & Margareta Grönberg

Authors
  1. E. C. J. Norrby
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  2. Pernilla Magnusson
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  3. L. -G. Falksveden
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  4. Margareta Grönberg
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Supported by grants from the Medical Research Fund of Karolinska Institutet and the Swedish Medical Research Council.

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Norrby, E.C.J., Magnusson, P., Falksveden, L.G. et al. Separation of measles virus components by equilibrium centrifugation in CsCl gradients. Archiv f Virusforschung 14, 462–473 (1964). https://doi.org/10.1007/BF01555078

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  • DOI: https://doi.org/10.1007/BF01555078

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