Cannons A C, Merrett M J
Eur J Biochem. 1984 Aug 1;142(3):597-602. doi: 10.1111/j.1432-1033.1984.tb08328.x.
Exposure of dark-grown Euglena to white or red light, but not blue light, produced a twofold increase in the specific activity of citrate synthase. A 400-fold purification of mitochondrial citrate synthase (subunit Mr = 44000) was achieved from cells of Euglena gracilis by affinity chromatography on ATP-activated agarose. Antisera, raised against the homogeneously pure enzyme, were used to demonstrate that the increase in citrate synthase activity on exposure of dark-grown cells to light resulted from an increase in citrate synthase protein. Anti-(citrate synthase) was used to detect precursor citrate synthase resulting from the translation of total polyadenylated RNA from Euglena in a cell-free rabbit reticulocyte lysate system. Citrate synthase mRNA was found to be present in cells at all stages of regreening. However, extraction and translation of polyadenylated RNA from free polysomes isolated from darkgrown and regreening cells demonstrated that appreciable translation of citrate synthase mRNA was only occurring in regreening cells.
将黑暗中生长的眼虫暴露于白光或红光而非蓝光下，会使柠檬酸合酶的比活性增加两倍。通过在ATP活化琼脂糖上进行亲和层析，从纤细眼虫细胞中实现了线粒体柠檬酸合酶（亚基分子量=44000）400倍的纯化。用针对同质纯酶产生的抗血清来证明，黑暗中生长的细胞暴露于光下时柠檬酸合酶活性的增加是由于柠檬酸合酶蛋白的增加。抗（柠檬酸合酶）用于在无细胞兔网织红细胞裂解物系统中检测由眼虫总多聚腺苷酸化RNA翻译产生的柠檬酸合酶前体。发现柠檬酸合酶mRNA在复绿的所有阶段的细胞中均存在。然而，从黑暗中生长和复绿细胞中分离的游离多核糖体中提取和翻译多聚腺苷酸化RNA表明，柠檬酸合酶mRNA的显著翻译仅发生在复绿细胞中。
