Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innsbruck, Austria.
Methods Mol Biol. 2024;2717:159-173. doi: 10.1007/978-1-0716-3453-0_10.
As allergies, especially those triggered by food, are becoming more and more prevalent, it is of increasing importance to fully understand the structures and dynamic behaviors of allergenic proteins along with their interactions with potential natural ligands. Therefore, we have established a solid routine to achieve structural characterization of food allergens, especially for birch pollen-related cross-reactive proteins from the class 10 of pathogenesis-related proteins (PR-10), by nuclear magnetic resonance (NMR) spectroscopy. Following expression of the desired allergen in Escherichia coli in isotope-labeled minimal media, the three-dimensional solution structures of these proteins can be determined, and insight into ligand binding mechanics and structural dynamic properties are accessible through NMR spin relaxation experiments.
随着过敏反应，特别是由食物引起的过敏反应变得越来越普遍，充分了解过敏原蛋白的结构和动态行为以及它们与潜在天然配体的相互作用变得越来越重要。因此，我们建立了一个可靠的常规程序，通过核磁共振（NMR）光谱法实现对食物过敏原的结构特征分析，特别是针对 10 类与桦树花粉相关的交叉反应蛋白（PR-10）的类过敏蛋白。通过在同位素标记的最小培养基中在大肠杆菌中表达所需的过敏原，这些蛋白质的三维溶液结构可以被确定，并且可以通过 NMR 自旋弛豫实验来深入了解配体结合力学和结构动态特性。
