Identification of rat brain polysomes synthesizing the brain specific enolase (14.3.2 protein), S100 protein and alpha and beta tubulin subunits |
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Authors: | A Marks M M Portier Y Zeitoun L Legault-Demare J Thibault N Lamande C Jeantet F Gros |
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Affiliation: | 1. Laboratoire de Biochimie Cellulaire, Collège de France, France;2. 11, Place Marcelin-Berthelot, 57231 Paris Cedex 05, France |
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Abstract: | Polysomes prepared from frozen rat brain powder were fractionated by centrifugation in a sucrose gradient. Individual fractions were used to program a reticulocyte lysate in a run-off reaction. The products of cell-free synthesis were assayed for the brain-specific enolase (14.3.2 protein) and S100 protein by immunoprecipitation with specific antisera and for tubulin by two-dimensional electrophoresis in polyacrylamide slab gels. The relative synthesis of these proteins by unfractionated free brain polysomes were 0.1 per cent, 0.05 per cent and 0.7 per cent respectively. After centrifugation in a sucrose gradient polysomes synthesizing S100 protein were separated from those synthesizing the other two markers. There was a threefold enrichment in the specific messenger RNA activity for each of the three proteins studied in their respective peak fractions of polysomes. |
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Keywords: | Polysomes de cerveaux de rat Protéine 14.32 Protéine S100 Tubuline Rat brain polysomes 14.3.2 protein S100 protein Tubulin TCA trichloroacetic acid SDS sodium dodecyl sulfate BTS Beckman tissue solubilizer |
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