Partial purification and characterization of messenger RNA coding 14-3-2 protein from rat brain.
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.
D000818:Animals; D001923:Brain Chemistry; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005559:Formamides; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D011061:Poly A / Q000302:isolation & purification; D014176:Protein Biosynthesis; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D012156:Reticulocytes / Q000378:metabolism
D005559:Formamides; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA