Carmo-Gonçalves Phelippe, Pinheiro Anderson S, Romão Luciana, Cortines Juliana, Follmer Cristian
Department of Physical Chemistry and.
Amyloid. 2014 Sep;21(3):163-74. doi: 10.3109/13506129.2014.912208. Epub 2014 Apr 30.
Oxidative stress and the formation of cytotoxic aggregates of the presynaptic protein α-synuclein (AS) are two important events associated with the pathogenesis of Parkinson's disease (PD) and several other neurodegenerative diseases. In this context, extensive efforts have been made to elucidate the molecular basis of the cytotoxic synergy between oxidative stress and AS aggregation. In this study, we demonstrate that the exposure of AS to oxidative stress induced by UV radiation (ASUV) blocks the protein fibrillation, leading to the formation of highly toxic fibril-incompetent oligomers. In addition, ASUV exhibited stronger anti-fibrillogenic properties than H2O2-treated AS, inhibiting the fibrillation of unmodified AS at notably low concentrations. Mass spectrometry indicated that Met5 oxidation to Met-sulfoxide was the only modification promoted by UV exposure, which is reinforced by NMR data indicating that Met5 is the only residue whose amide resonance completely disappeared from the (1)H-(15)N HSQC spectrum after UV exposure. This result is supported by previous data that indicate that C-terminal Met residues (Met116 and Met127) and N-terminal Met1 are less susceptible to oxidation than Met5 because of the residual structure of the disordered AS monomer. Overall, our findings suggest that specific oxidation of Met5 might be sufficient to promote the formation of highly neurotoxic oligomers of AS.
氧化应激和突触前蛋白α-突触核蛋白(AS)细胞毒性聚集体的形成是与帕金森病(PD)及其他几种神经退行性疾病发病机制相关的两个重要事件。在此背景下,人们已做出大量努力来阐明氧化应激与AS聚集之间细胞毒性协同作用的分子基础。在本研究中,我们证明将AS暴露于紫外线辐射诱导的氧化应激(ASUV)会阻止蛋白质纤维化,导致形成高毒性的无纤维化能力的寡聚体。此外,ASUV比经H2O2处理的AS表现出更强的抗纤维化特性,在极低浓度下就能抑制未修饰AS的纤维化。质谱分析表明,紫外线暴露促进的唯一修饰是Met5氧化为甲硫氨酸亚砜,核磁共振数据进一步证实了这一点,该数据表明Met5是紫外线暴露后(1)H-(15)N HSQC谱中酰胺共振完全消失的唯一残基。先前的数据支持了这一结果,这些数据表明由于无序AS单体的残余结构,C端的Met残基(Met116和Met127)和N端的Met1比Met5更不易被氧化。总体而言,我们的研究结果表明,Met5的特定氧化可能足以促进AS形成高度神经毒性的寡聚体。