CHAUVEAU J, MOULE Y, ROUILLER C, SCHNEEBELI J
J Cell Biol. 1962 Jan;12(1):17-29. doi: 10.1083/jcb.12.1.17.
Microsomes, isolated from rat liver homogenate in 0.88 M sucrose, have been fractionated by differential centrifugation. The 2nd microsomal fraction, sedimented between 60 minutes at 105,000 g and 3 hours at 145,000 g, consists mainly of smooth vesicles, free ribosomes, and ferritin. By utilizing the differences in density existing between the membranes and the granular elements it has been possible to separate the smooth membranes from the free ribosomes and ferritin. The procedure is to resuspend the 2nd microsomal fraction in a sucrose solution of 1.21 or 1.25 density and centrifuge it at 145,000 g for 20 or 40 hours. A centripetal migration of membranes and a centrifugal sedimentation of granular elements are obtained. Phospholipids, as well as the enzymatic activities DPNH-cytochrome c reductase, glucose-6-phosphatase and esterase are localized in the membranes. The free ribosomes have been purified by washing. A concentration of 200 microg RNA per mg nitrogen has been reached. RNA is also present in the membranes. These results are discussed in relation to current views on microsomal structure and chemistry.
从大鼠肝脏匀浆在0.88 M蔗糖中分离得到的微粒体,已通过差速离心进行了分级分离。第二个微粒体级分,在105,000 g下离心60分钟至145,000 g下离心3小时之间沉淀,主要由光滑囊泡、游离核糖体和铁蛋白组成。利用膜与颗粒成分之间存在的密度差异,已能够将光滑膜与游离核糖体和铁蛋白分离。该步骤是将第二个微粒体级分重悬于密度为1.21或1.25的蔗糖溶液中,并在145,000 g下离心20或40小时。得到了膜的向心迁移和颗粒成分的离心沉淀。磷脂以及酶活性二磷酸吡啶核苷酸 - 细胞色素c还原酶、葡萄糖 - 6 - 磷酸酶和酯酶定位于膜中。游离核糖体已通过洗涤纯化。已达到每毫克氮200微克RNA的浓度。RNA也存在于膜中。结合当前关于微粒体结构和化学的观点对这些结果进行了讨论。