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In vitro synthesis of two brain-specific proteins (S100 and 14-3-2) by polyribosomes from rat brain

Archives of Biochemistry and Biophysics, ISSN: 0003-9861, Vol: 155, Issue: 1, Page: 58-69
1973
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Free and membrane-bound polyribosomes were prepared from whole brain, cortex, and hindbrain-medullary white matter of young adult rats. In a homologous cell-free system, two brain-specific acidic proteins (S100 and 14-3-2) were identified in the soluble proteins released from the polyribosomes during incubation for 1 hr at 37°C. Both S100 and 14-3-2 proteins were found to be synthesized primarily by free polyribosomes. De novo synthesis of the 14-3-2 protein was achieved in a reconstituted cerebral cell-free system containing polysome-derived DNA-like RNA and 40S+60S subunits. The radioactively labeled S100 and 14-3-2 proteins were identified by immunoprecipitation with specific antisera and sodium dodecyl sulfate polyacrylamide gel electrophoresis. No difference was found in the synthesis of these brain-specific proteins when a hepatic supernatant enzyme fraction was used in the cell-free system containing cerebral polyribosomes. Furthermore, in vitro synthesis of S100 and 14-3-2 proteins did not occur with liver polysomes when a supernatant enzyme fraction from either liver or brain was used.

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