Fausnaugh-Pollitt J, Thevenon G, Janis L, Regnier F E
Department of Chemistry, Purdue University, West Lafayette, IN 47907.
J Chromatogr. 1988 Jun 29;443:221-8. doi: 10.1016/s0021-9673(00)94795-2.
There are seven avian lysozyme variants of nearly identical three-dimensional structure which have amino acid substitutions broadly distributed on their surface. By using these protein variants, it was possible to study the relationship between protein structure and chromatographic retention. It was determined that according to the mode of separation various regions of the proteins surface determine chromatographic retention. At one extreme, immunosorbents targeted a very small region on the protein surface. Hydrophobic interaction chromatography was an intermediate case in which one surface domain of the lysozymes controlled chromatographic behavior. At the opposite extreme, cation-exchange columns probed most of the protein surface. It was concluded that identification of random variations in protein structure will be most successfully detected by a separation mode that broadly targets the surface of a protein.
有七种三维结构几乎相同的鸟类溶菌酶变体，其氨基酸取代广泛分布在它们的表面。通过使用这些蛋白质变体，研究蛋白质结构与色谱保留之间的关系成为可能。结果确定，根据分离模式，蛋白质表面的不同区域决定了色谱保留。在一个极端情况下，免疫吸附剂靶向蛋白质表面的一个非常小的区域。疏水相互作用色谱是一种中间情况，其中溶菌酶的一个表面结构域控制色谱行为。在另一个极端，阳离子交换柱探测了大部分蛋白质表面。得出的结论是，通过广泛靶向蛋白质表面的分离模式将最成功地检测到蛋白质结构中的随机变异。
