Norling B, Zak E, Andersson B, Pakrasi H
Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden.
FEBS Lett. 1998 Oct 2;436(2):189-92. doi: 10.1016/s0014-5793(98)01123-5.
Aqueous polymer two-phase partitioning in combination with sucrose density centrifugation offered, for the first time, a 2D-separation method for the isolation of pure plasma and thylakoid membranes from the cyanobacterium Synechocystis 6803 without any cross-contaminations. The purity of the membrane fractions was verified by immunoblot analysis using antibodies against membrane-specific marker proteins. As an initiation of a proteomics project, two prominent proteins, which were observed only in the plasma membrane (Slr1513, a hypothetical protein, and HofG, a general secretion pathway protein), or in the thylakoid membrane (PsaE, a photosystem I protein, and NdhH, a subunit of NADH dehydrogenase), were identified.
水性聚合物双相分配结合蔗糖密度离心首次提供了一种二维分离方法，用于从集胞藻6803中分离纯质膜和类囊体膜，且无任何交叉污染。通过使用针对膜特异性标记蛋白的抗体进行免疫印迹分析，验证了膜组分的纯度。作为蛋白质组学项目的开端，鉴定出了两种显著的蛋白质，一种仅在质膜中观察到（Slr1513，一种假定蛋白，和HofG，一种通用分泌途径蛋白），另一种仅在类囊体膜中观察到（PsaE，一种光系统I蛋白，和NdhH，NADH脱氢酶的一个亚基）。
