LABELLA F S, BROWN J H
J Biophys Biochem Cytol. 1959 Jan 25;5(1):17-24. doi: 10.1083/jcb.5.1.17.
Fresh anterior pituitary glands from beef and pig were separated by differential centrifugation into subcellular fractions. Nuclei and debris were obtained at 700 g for 15 minutes, secretory granules at 7000 g for 20 minutes, mitochondria at 34,000 g for 15 minutes, and microsomes at 78,000 g for 3 hours. Electron micrographs were taken of the individual fractions. Each fraction was analyzed for nitrogen, pentosenucleic acid (PNA), and phospholipide. Beef and pig anterior lobes were quite similar in their intracellular composition as seen in the subcellular fractions. Succinic dehydrogenase was localized in mitochondria, while alkaline phosphatase was concentrated in the microsomes. A proteinase with pH optimum at 8.2 was exclusively localized. in microsomal and supernatant fractions. Acid phosphatase, acid ribonuclease, and acid proteinase were distributed among the subcellular fractions in another pattern, indicating the presence of a particle type distinct from mitochondria and microsomes. The distribution of cytoplasmic PNA paralleled that of alkaline phosphatase.
通过差速离心法将新鲜的牛和猪垂体前叶分离成亚细胞组分。在700g下离心15分钟获得细胞核和碎片,在7000g下离心20分钟获得分泌颗粒,在34000g下离心15分钟获得线粒体,在78000g下离心3小时获得微粒体。对各个组分拍摄电子显微镜照片。分析每个组分中的氮、戊糖核酸(PNA)和磷脂。从亚细胞组分来看,牛和猪的垂体前叶在细胞内组成上非常相似。琥珀酸脱氢酶定位于线粒体中,而碱性磷酸酶集中在微粒体中。一种最适pH为8.2的蛋白酶仅定位于微粒体和上清液组分中。酸性磷酸酶、酸性核糖核酸酶和酸性蛋白酶以另一种模式分布于亚细胞组分中,表明存在一种不同于线粒体和微粒体的颗粒类型。细胞质PNA的分布与碱性磷酸酶的分布平行。