Widell S, Lundborg T, Larsson C
Department of Plant Physiology, University of Lund, S-220 07 Lund, Sweden.
Plant Physiol. 1982 Nov;70(5):1429-35. doi: 10.1104/pp.70.5.1429.
Presumptive plasma membrane fractions have been prepared from oat (Avena sativa L. cv. Brighton) roots and shoots, respectively, by partition of microsomal fractions in a dextran-polyethylene glycol two-phase system. The plasma membranes had a high affinity for the polyethylene glycol-rich upper phase, whereas membranes from mitochondria and other organelles partitioned in the dextran-rich lower phase or at the interface. Thus, relatively pure plasma membranes were obtained by only two partition steps, and within 3 hours from homogenization of the material.The plasma membranes from both organs were enriched in K(+)-stimulated Mg(2+)-dependent ATPase and glucan synthetase II, two tentative markers for the plant plasma membrane. Silicotungstic acid, an indicative stain for the plasma membrane, stained the vesicles recovered from the upper phase, but failed to stain the membranes partitioning in the lower phase or at the interface.The plasma membranes were also enriched in a light-reducible b-cytochrome. This b-cytochrome can be measured by its light-induced absorbance change and may serve as a marker for the plant plasma membrane.
通过在葡聚糖 - 聚乙二醇两相系统中对微粒体部分进行分配,分别从燕麦( Avena sativa L. cv. Brighton)的根和芽中制备了推定的质膜部分。质膜对富含聚乙二醇的上相具有高亲和力,而线粒体和其他细胞器的膜则分配在富含葡聚糖的下相或界面处。因此,仅通过两个分配步骤,并且在材料匀浆后的3小时内就获得了相对纯的质膜。来自两个器官的质膜富含K(+)刺激的Mg(2+)依赖性ATP酶和葡聚糖合成酶II,这是植物质膜的两个暂定标记物。硅钨酸是质膜的指示性染色剂,它能使从上相中回收的囊泡染色,但不能使分配在下相或界面处的膜染色。质膜中还富含一种可光还原的b - 细胞色素。这种b - 细胞色素可以通过其光诱导的吸光度变化来测量,并且可以作为植物质膜的标记物。