Sakiyama S, Akiba N, Fujimura S
Cancer Res. 1979 Feb;39(2 Pt 1):502-6.
The messenger activity for fructose 1,6-bisphosphate aldolase (EC4.1.2.13) (aldolase) A isozyme has been characterized in the polysome- or the messenger RNA-directed, protein-synthesizing system using the pH 5 fraction of rat liver or wheat germ extracts, respectively. The subunit of aldolase A synthesized in vitro was detected by immunoprecipitation with anti-aldolase A antibody raised in chickens followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The synthesis of the enzyme depended on the addition of polysomes or polyadenylate-containing RNA of rat ascites hepatoma AH 7974 cells which show a complete shift of aldolase isozyme to type A, whereas polysomes of adult rat liver were inactive. The messenger activity for aldolase A was present exclusively on free polysomes but absent on membrane-bound polysomes and in the soluble supernatant fraction of AH 7974 cells. The size of aldolase A messenger RNA determined by formamide-containing sucrose density gradient centrifugation was approximately 5.8 X 10(5) daltons corresponding to 1650 nucleotides. Taking into account the number of amino acid residues in the aldolase A subunit, approximately 400 nucleotides correspond to the noncoding region of aldolase A messenger RNA.
已分别使用大鼠肝脏的pH 5组分或小麦胚芽提取物，在多核糖体或信使核糖核酸指导的蛋白质合成系统中对1,6 - 二磷酸果糖醛缩酶（EC4.1.2.13）（醛缩酶）A同工酶的信使活性进行了表征。体外合成的醛缩酶A亚基通过用鸡产生的抗醛缩酶A抗体进行免疫沉淀，然后进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳来检测。该酶的合成取决于添加大鼠腹水肝癌AH 7974细胞的多核糖体或含聚腺苷酸的RNA，这些细胞的醛缩酶同工酶完全转变为A型，而成年大鼠肝脏的多核糖体则无活性。醛缩酶A的信使活性仅存在于游离多核糖体上，而在AH 7974细胞的膜结合多核糖体和可溶性上清液组分中不存在。通过含甲酰胺的蔗糖密度梯度离心法测定的醛缩酶A信使核糖核酸的大小约为5.8×10⁵道尔顿，相当于1650个核苷酸。考虑到醛缩酶A亚基中的氨基酸残基数，约400个核苷酸对应于醛缩酶A信使核糖核酸的非编码区。
