Sato S, Asahi T
Laboratory of Biochemistry, Faculty of Agriculture, Nagoya University, Chikusa, Nagoya 464, Japan.
Plant Physiol. 1975 Dec;56(6):816-20. doi: 10.1104/pp.56.6.816.
An attempt to isolate intact mitochondria from dry pea seeds (Pisum sativum var. Alaska) ended in failure. Cytochrome oxidase in crude mitochondrial fraction from dry seeds was separated into three fractions by sucrose density gradient centrifugation. Two of the fractions contained malate dehydrogenase, whereas the other did not. Equilibrium centrifugation of mitochondrial membrane on sucrose gradients revealed that the membrane from the fraction without malate dehydrogenase was lighter than that from the others. Differences were observed in relative content of phospholipid to protein and in polypeptide composition analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis among the membranes from three fractions and imbibed cotyledons. Membrane from the fraction without malate dehydrogenase was rich in phospholipid and lacking in polypeptides with relatively high molecular weights as compared with that from others. During imbibition, the fraction without malate dehydrogenase and one of the other two disappeared rapidly after a lag phase lasting for at least 1 hour. Concomitantly, active and stable mitochondria increased in the cotyledons. The results were interpreted to indicate that there were at least three types of mitochondria in dry seeds, the membranes of which differed in their biochemical properties, and that the mitochondria became active and stable through assembly of protein into the membranes during imbibition.
尝试从干豌豆种子（阿拉斯加豌豆品种）中分离完整的线粒体，但以失败告终。通过蔗糖密度梯度离心法，将干种子粗线粒体组分中的细胞色素氧化酶分离成三个组分。其中两个组分含有苹果酸脱氢酶，而另一个组分则没有。在线粒体膜的蔗糖梯度平衡离心中发现，不含苹果酸脱氢酶的组分的膜比其他组分的膜轻。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析，在三个组分和吸胀子叶的膜中观察到磷脂与蛋白质的相对含量以及多肽组成存在差异。与其他组分相比，不含苹果酸脱氢酶的组分的膜富含磷脂，且缺乏相对高分子量的多肽。在吸胀过程中，不含苹果酸脱氢酶的组分和另外两个组分中的一个在持续至少1小时的滞后期后迅速消失。与此同时，子叶中活跃且稳定的线粒体增加。结果表明，干种子中至少存在三种类型的线粒体，其膜的生化特性不同，并且在吸胀过程中，通过蛋白质组装到膜中，线粒体变得活跃且稳定。
