Ackers G K, Smith F R
Biophys J. 1986 Jan;49(1):155-65. doi: 10.1016/S0006-3495(86)83631-1.
Site-specific structural modification is a powerful tool for studying functional mechanisms in proteins where the structures may be manipulated by direct chemical modification, by selection of naturally-occurring mutants, or by site-directed mutagenesis. Here, we present a general strategy for such studies, which we term "mapping by structure-function perturbation." A series of functional perturbations (i.e., deviations of functional behavior from that of the native protein) are mapped against the structural locations of the modified sites, obtained over a range of locations. The modifications are treated as arbitrary perturbations of structure at specific locations, in contrast to the conventional approach of trying to interpret their local stereochemistry. The map yields information on structural locations of functional events and pathways of coupling within protein assemblies. We have applied this approach to the ligand-linked subunit assembly of human hemoglobin, using both chemically-modified heme sites (CN-met), and amino acid residues altered by mutation and chemical modification.
位点特异性结构修饰是研究蛋白质功能机制的有力工具，在蛋白质中，结构可以通过直接化学修饰、选择天然存在的突变体或定点诱变来进行操控。在此，我们提出了一种用于此类研究的通用策略，我们称之为“结构-功能扰动图谱法”。一系列功能扰动（即功能行为与天然蛋白质的偏差）与在一系列位置上获得的修饰位点的结构位置相对应。与试图解释其局部立体化学的传统方法不同，这些修饰被视为特定位置结构的任意扰动。该图谱可提供有关功能事件的结构位置以及蛋白质组装体内偶联途径的信息。我们已将此方法应用于人血红蛋白的配体连接亚基组装，使用了化学修饰的血红素位点（氰化高铁血红蛋白）以及通过突变和化学修饰改变的氨基酸残基。
