Kratzin H D, Kruse T, Maywald F, Thinnes F P, Götz H, Egert G, Pauly E, Friedrich J, Yang C Y, Wernet P
J Chromatogr. 1984 Aug 3;297:1-11. doi: 10.1016/s0021-9673(01)89023-3.
The amino acid sequence of the human HLA-DR2 class II antigen was determined with the aid of high-performance liquid chromatography. By applying exclusively Hypersil-ODS as the stationary phase in combination with three buffer systems, the amino acid sequences of the extracellular parts of the DR alpha- and DR beta-chains could be established. Moreover, the sensitivity of the procedure allowed the determination of the primary structures of additional class II polypeptides isolated together with the DR2 alpha- and the DR2 beta-chains. Significant insight into the heterogeneity of class II molecules was thereby obtained. In addition, a simple, rapid and selective method of general applicability is described for the isolation of membrane fragments of integral membrane proteins. Using Sep-Pak cartridges with a mobile phase containing 60% formic acid and varying 1-propanol concentrations, the highly hydrophobic peptides can selectively be separated from the hydrophilic fragments.
借助高效液相色谱法测定了人类HLA - DR2Ⅱ类抗原的氨基酸序列。通过仅使用Hypersil - ODS作为固定相并结合三种缓冲系统，得以确定DRα链和DRβ链细胞外部分的氨基酸序列。此外，该方法的灵敏度使得能够确定与DR2α链和DR2β链一起分离出的其他Ⅱ类多肽的一级结构。由此对Ⅱ类分子的异质性有了重要认识。此外，还描述了一种简单、快速且具有普遍适用性的选择性方法，用于分离整合膜蛋白的膜片段。使用含有60%甲酸和不同浓度1 - 丙醇的流动相的Sep - Pak柱，可以将高度疏水的肽段与亲水性片段选择性地分离。
