Kamp R M, Bosserhoff A, Kamp D, Wittmann-Liebold B
J Chromatogr. 1984 Dec 28;317:181-92. doi: 10.1016/s0021-9673(01)91658-9.
The ribosomal proteins from Escherichia coli, Bacillus stearothermophilus and Methanococcus vannielii were separated by size-exclusion, ion-exchange and reversed-phase high-performance liquid chromatography (HPLC), employing new column materials, different gradient systems, and preparative columns, respectively. The purity of the isolated proteins was analysed by one- and two-dimensional gel electrophoresis and by direct micro-sequencing. The separation of ribosomal proteins could be improved by employing propanol gradients in combination with Vydac reversed-phase columns. From the E. coli ribosome, fifteen S and twenty-three L proteins were isolated in sequencer purity by this method. In addition, ion-exchange HPLC was proven to be useful for isolating ribosomal proteins under native conditions: six S proteins and sixteen L proteins from E. coli could be purified. Some of these proteins were not isolated by the reversed-phase procedures, e.g. proteins L9, L14 and L21.
分别采用新型柱材料、不同梯度系统和制备柱，通过尺寸排阻、离子交换和反相高效液相色谱（HPLC）对来自大肠杆菌、嗜热脂肪芽孢杆菌和万氏甲烷球菌的核糖体蛋白进行分离。通过一维和二维凝胶电泳以及直接微量测序分析分离得到的蛋白质的纯度。结合使用丙醇梯度和Vydac反相柱可改善核糖体蛋白的分离效果。用这种方法从大肠杆菌核糖体中分离出了序列分析纯的15种S蛋白和23种L蛋白。此外，离子交换HPLC被证明可用于在天然条件下分离核糖体蛋白：可纯化出大肠杆菌的6种S蛋白和16种L蛋白。其中一些蛋白质不能通过反相方法分离，例如L9、L14和L21蛋白。
