Noguchi T, Tanaka T
J Biol Chem. 1982 Feb 10;257(3):1110-3.
The question of whether the M1- and M2-type isozymes of pyruvate kinase are synthesized by a common mRNA was examined by isolating total RNA from rat skeletal muscle and AH-130 Yoshida ascites hepatoma cells, which express the M1- and M2-type isozymes, respectively, and translating the preparations in a rabbit reticulocyte lysate system. The newly synthesized pyruvate kinase subunits were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing under the denaturing condition. The pyruvate kinase subunit synthesized from hepatoma cell RNA had a slightly larger molecular weight and higher pI value than the subunit from muscle RNA. When a mixture of the two RNAs was added to the lysate, the same result was obtained. Thus, it is concluded that the M1- and M2-type isozymes are translated from different messenger RNAs.
通过从大鼠骨骼肌和AH-130吉田腹水肝癌细胞中分离总RNA来研究丙酮酸激酶的M1型和M2型同工酶是否由共同的mRNA合成,这两种细胞分别表达M1型和M2型同工酶,并在兔网织红细胞裂解物系统中翻译这些制剂。在变性条件下,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和等电聚焦比较新合成的丙酮酸激酶亚基。从肝癌细胞RNA合成的丙酮酸激酶亚基的分子量略大于肌肉RNA亚基,且pI值更高。当将两种RNA的混合物添加到裂解物中时,得到了相同的结果。因此,可以得出结论,M1型和M2型同工酶是由不同的信使RNA翻译而来的。