Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331.
Proc Natl Acad Sci U S A. 1984 Aug;81(15):4804-8. doi: 10.1073/pnas.81.15.4804.
An isoelectric focusing method is described that discriminates plant cell organelle populations on the basis of surface charge. The isoelectric points (pI values) of the Golgi apparatus, the mitochondria, and putative plasma membrane from etiolated pea stem cells are reported. The pI of the pea Golgi apparatus is similar to that of the (35)SO(4)-labeled membranes of developing Fucus embryos. The pI of Fucus(35)SO(4)-labeled membranes depends on whether the membranes are being polarly transported to the growing tip or randomly transported to the entire periphery of the zygote. Those subject to polar tip transport have a significantly greater negative surface charge than those being randomly transported. The implication of this result toward an understanding of the localization mechanism is discussed. The method is also capable of subfractionating glucan synthetase II-containing membranes (putative plasma membrane) from peas. The pI of putative plasma membrane from a pea stem homogenate is similar to the pI of the plasma membrane of whole protoplasts of Catharanthus leaf cells. Isoelectric focusing appears to be a useful technique to discriminate membranes and, hence, to provide new information and approaches to study cellular and developmental phenomena.
描述了一种等电聚焦方法,该方法基于表面电荷区分植物细胞细胞器群体。报告了黄化豌豆茎细胞的高尔基器、线粒体和假定质膜的等电点(pI 值)。豌豆高尔基器的 pI 与发育中的石莼胚胎的(35)SO4 标记膜相似。石莼(35)SO4 标记膜的 pI 取决于膜是被极性运输到生长尖端还是随机运输到合子的整个周边。那些受极性尖端运输的膜比随机运输的膜具有显著更大的负表面电荷。讨论了这一结果对理解定位机制的意义。该方法还能够从小豆中分离含葡聚糖合成酶 II 的膜(假定质膜)。豌豆茎匀浆中假定质膜的 pI 与长春花叶片细胞原生质体完整质膜的 pI 相似。等电聚焦似乎是一种有用的技术,可以区分膜,从而为研究细胞和发育现象提供新的信息和方法。