Barbieri Letizia, Luchinat Enrico
Magnetic Resonance Center, University of Florence, Via Luigi Sacconi 6, 50019, Sesto Fiorentino, Italy.
Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine, Via Luigi Sacconi 6, Sesto Fiorentino, Italy.
Biomol NMR Assign. 2019 Oct;13(2):371-376. doi: 10.1007/s12104-019-09908-8. Epub 2019 Aug 3.
DJ-1 is a highly conserved soluble protein that is associated to several cellular pathways. In humans, DJ-1 has been implicated in several pathologies such as cancer, Parkinson's disease and amyotrophic lateral sclerosis. Several roles have been attributed to DJ-1, including defense against oxidative stress, chaperone activity and proteasome regulation. The recent finding that DJ-1 acts as a protein and DNA deglycase further confirms the protective function of DJ-1 and suggests a common mechanism of action in the various pathways in which DJ-1 is involved. Cysteine 106, located in the putative active site of DJ-1, is critical for the biological activity of DJ-1 and is easily oxidized to cysteine-sulfinate. While such oxidation modulates DJ-1 activity, the underlying molecular mechanism has not yet been elucidated. Cysteine oxidation does not perturb the protein structure, therefore changes in protein dynamics in solution could modulate its function. Here, we report a revised and completed (98%) backbone assignment of reduced DJ-1, together with the backbone assignment of oxidized DJ-1. Chemical shift perturbation is observed in several regions across the sequence, while no changes in secondary structure are observed. These data will provide the starting point for further characterization of the changes in the backbone dynamics of DJ-1 upon oxidation in solution at physiological temperature.
DJ-1是一种高度保守的可溶性蛋白质,与多种细胞途径相关。在人类中,DJ-1与多种疾病有关,如癌症、帕金森病和肌萎缩侧索硬化症。DJ-1具有多种作用,包括抗氧化应激、伴侣活性和蛋白酶体调节。最近发现DJ-1可作为一种蛋白质和DNA去糖基化酶,这进一步证实了DJ-1的保护功能,并提示了DJ-1所涉及的各种途径中的共同作用机制。位于DJ-1假定活性位点的半胱氨酸106对DJ-1的生物学活性至关重要,并且很容易被氧化为半胱氨酸亚磺酸盐。虽然这种氧化调节DJ-1的活性,但其潜在的分子机制尚未阐明。半胱氨酸氧化不会干扰蛋白质结构,因此溶液中蛋白质动力学的变化可能会调节其功能。在这里,我们报告了还原型DJ-1的修订和完整(98%)的主链归属,以及氧化型DJ-1的主链归属。在整个序列的几个区域观察到化学位移扰动,而二级结构没有变化。这些数据将为进一步表征生理温度下溶液中DJ-1氧化后主链动力学变化提供起点。