Quasi-elastic light scattering of Artemia ribosomes sedimented in a CsCl density gradient |
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Authors: | Paul Nieuwenhuysen Julius Clauwaert |
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Affiliation: | Department of Cell Biology, University of Antwerp, 2610 Wilrijk, Belgium |
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Abstract: | It is impossible to measure the diffusion coefficient of macromolecules directly and accurately by quasi—elastic light scattering, when aggregates cannot be eliminated from the solutions to be investigated. Nevertheless, a simple method can be applied to overcome this problem in many cases. Aggregates are separated from the monomeric macromolecules by rate-zonal sedimentation in a CsCl density gradient in a transparent centrifugation tube; the monomers are then located by laser light scattering intensity measurements; photon correlation spectroscopy of the scattered light finally yields their diffusion coefficient. The viscosity of aqueous CsCl solutions at different temperatures and concentrations allows a good separation by centrifugation and a low uncertainty in the reduction of the measured diffusion coefficient to standard conditions.The application of the method to eukaryotic large ribosomal subunits is described as an example. |
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Keywords: | quasi-elastic light scattering photon correlation spectroscopy density-gradient centrifugation diffusion coefficient ribosomes Hepes N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid |
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