Girod Marion, Enjalbert Quentin, Brunet Claire, Antoine Rodolphe, Lemoine Jérôme, Lukac Iva, Radman Miroslav, Krisko Anita, Dugourd Philippe
Université de Lyon, 69622, Lyon, France; Institut des Sciences Analytiques, UMR 5280, CNRS, Université Claude Bernard Lyon 1, Lyon, France.
Université de Lyon, 69622, Lyon, France; Institut Lumière Matière, UMR 5306, CNRS, Université Claude Bernard Lyon 1, Lyon, France.
PLoS One. 2014 Jul 7;9(7):e101642. doi: 10.1371/journal.pone.0101642. eCollection 2014.
Accumulation of oxidative damage in proteins correlates with aging since it can cause irreversible and progressive degeneration of almost all cellular functions. Apparently, native protein structures have evolved intrinsic resistance to oxidation since perfectly folded proteins are, by large most robust. Here we explore the structural basis of protein resistance to radiation-induced oxidation using chicken egg white lysozyme in the native and misfolded form. We study the differential resistance to oxidative damage of six different parts of native and misfolded lysozyme by a targeted tandem/mass spectrometry approach of its tryptic fragments. The decay of the amount of each lysozyme fragment with increasing radiation dose is found to be a two steps process, characterized by a double exponential evolution of their amounts: the first one can be largely attributed to oxidation of specific amino acids, while the second one corresponds to further degradation of the protein. By correlating these results to the structural parameters computed from molecular dynamics (MD) simulations, we find the protein parts with increased root-mean-square deviation (RMSD) to be more susceptible to modifications. In addition, involvement of amino acid side-chains in hydrogen bonds has a protective effect against oxidation Increased exposure to solvent of individual amino acid side chains correlates with high susceptibility to oxidative and other modifications like side chain fragmentation. Generally, while none of the structural parameters alone can account for the fate of peptides during radiation, together they provide an insight into the relationship between protein structure and susceptibility to oxidation.
蛋白质中氧化损伤的积累与衰老相关,因为它会导致几乎所有细胞功能发生不可逆的渐进性退化。显然,天然蛋白质结构已经进化出对氧化的内在抗性,因为绝大多数折叠完美的蛋白质最为稳定。在这里,我们使用天然和错误折叠形式的鸡卵清溶菌酶来探索蛋白质对辐射诱导氧化的抗性的结构基础。我们通过对其胰蛋白酶片段的靶向串联/质谱方法研究了天然和错误折叠溶菌酶六个不同部分对氧化损伤的差异抗性。发现随着辐射剂量增加,每个溶菌酶片段的量的衰减是一个两步过程,其特征在于它们的量呈双指数演变:第一步很大程度上可归因于特定氨基酸的氧化,而第二步对应于蛋白质的进一步降解。通过将这些结果与从分子动力学(MD)模拟计算得到的结构参数相关联,我们发现均方根偏差(RMSD)增加的蛋白质部分更容易发生修饰。此外,氨基酸侧链参与氢键对氧化具有保护作用。单个氨基酸侧链暴露于溶剂的增加与对氧化和其他修饰(如侧链断裂)的高敏感性相关。一般来说,虽然没有一个结构参数单独能够解释辐射过程中肽段的命运,但它们共同提供了对蛋白质结构与氧化敏感性之间关系的深入了解。