Wassler M, Jonasson I, Persson R, Fries E
Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Biochem J. 1987 Oct 15;247(2):407-15. doi: 10.1042/bj2470407.
Monolayer cultures of rat hepatocytes were treated with increasing concentrations of saponin (prepared from Gypsophila plants) for 30 min at 6 degrees C. Differential permeabilization of the intracellular membranes could be demonstrated: at 0.040 mg of saponin/ml the plasma membrane was permeabilized, as assessed by the release of 50% of the total cellular amount of lactate dehydrogenase, and at 0.20 mg/ml the endoplasmic reticulum was permeabilized, as measured by the release of 50% of pulse-35S-labelled albumin. The Golgi complex was permeabilized at an intermediate saponin concentration, as indicated by the release of homogeneously 35S-labelled albumin; about half the intracellular albumin is located in this organelle. At 1.0 up to 5.0 mg of saponin/ml 90-95% of the radioactively labelled albumin was released. Even at 5.0 mg/ml less than 10% of the membrane of the endoplasmic reticulum was solubilized, as judged by the degree of release of a membrane-bound enzyme specific for this organelle. These results demonstrate the usefulness of saponin as a tool for investigating the interior of different intracellular compartments.
将大鼠肝细胞单层培养物在6℃下用浓度不断增加的皂角苷（从丝石竹属植物中制备）处理30分钟。可证明细胞内膜存在差异通透性：当皂角苷浓度为0.040毫克/毫升时，质膜通透性增加，这可通过乳酸脱氢酶总细胞量释放50%来评估；当浓度为0.20毫克/毫升时，内质网通透性增加，这通过脉冲-35S标记白蛋白释放50%来测定。在中间皂角苷浓度下高尔基体通透性增加，这由均匀35S标记白蛋白的释放表明；约一半细胞内白蛋白位于该细胞器中。在皂角苷浓度为1.0至5.0毫克/毫升时，90 - 95%的放射性标记白蛋白被释放。即使在5.0毫克/毫升时，根据该细胞器特异性膜结合酶的释放程度判断，内质网的膜溶解不到10%。这些结果证明了皂角苷作为研究不同细胞内区室内部工具的有用性。
