Assessment of protein structure and interaction is crucial for understanding protein structure/function relationships. Compared to high-resolution structural tools, including X-ray crystallography, nuclear magnetic resonance (NMR), and cryo-EM, and traditional low-resolution methods, such as circular dichroism, UV–vis, and florescence spectroscopy, mass spectrometry (MS)-based protein footprinting affords medium-to-high resolution (i.e., regional and residue-specific insights) by taking advantage of proteomics methods focused on the primary structure. The methodology relies on “painting” the reactive and solvent-exposed amino acid residues with chemical tags and using the pattern of modifications as footprints from analysis by bottom-up MS-based proteomics to deduce protein higher order structures. The outcome can refer to proteins in solution or even in cells and is complementary to those of X-ray crystallography and NMR. It is particularly useful in mapping protein–ligand interfaces and conformational changes resulting from ligand binding, mutation, and aggregation.

中文翻译：

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基于质谱的蛋白质快速光化学氧化（FPOP），用于高级结构表征

蛋白质结构和相互作用的评估对于理解蛋白质结构/功能关系至关重要。与高分辨率结构工具（包括X射线晶体学，核磁共振（NMR）和冷冻电磁场）以及传统的低分辨率方法（例如，圆二色性，紫外可见光谱和荧光光谱，质谱（MS））相比基于）的蛋白质足迹通过利用专注于一级结构的蛋白质组学方法，提供了中到高分辨率（即区域性和残基特异性的见解）。该方法依赖于用化学标签“涂装”反应性和溶剂暴露的氨基酸残基，并使用修饰的模式作为足迹，这些足迹是基于自下而上的基于MS的蛋白质组学分析得出的蛋白质高阶结构。结果可能涉及溶液中甚至是细胞中的蛋白质，并且与X射线晶体学和NMR的蛋白质互补。它在绘制蛋白质-配体界面以及由于配体结合，突变和聚集而引起的构象变化时特别有用。