Partial Purification and Characterization of Messenger RNA Coding 14‐3‐2 Protein from Rat Brain

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Abstract
14‐3‐2 Protein (neuron‐specific enolase) is a neuron‐specific protein. Using a reticulocyte lysate cell‐free system for translation of 14‐3‐2 protein mRNA, we have partially purified this mRNA by several procedures, including formamide sucrose density centrifugation, formamide polyacrylamide gel electrophoresis (PAGE) and polyuridylic acid (poly(U))‐Sepharose affinity chromatography. Using mRNA obtained by these procedures, we could increase the translation ratio of 14‐3‐2 protein synthesized/total soluble protein synthesized to 7.31%. The overall purification was 37.8‐fold. The size of 14‐3‐2 protein mRNA appears to be about 19–20S, because translation activity of mRNA obtained by sucrose density gradient centrifugation or formamide PAGE was the most active in this RNA size.