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

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, it was increased partially 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, the translation ratio of 14-3-2 protein synthesized/total soluble protein synthesized could be increased to 7.31%. The overall purification was 37.8-fold. The size of 14-3-2 protein mRNA appears to be .apprx. 19-20S, because translation acitvity of mRNA obtained by sucrose density gradient centrifugation or formamide PAGE was the most active in this RNA size.