METALLATION LEVELS OF PROTEIN AGGREGATES FORMED IN AMYOTROPHIC LATERAL SCLEROSIS

这个子项目是利用资源的许多研究子项目之一 由NIH/NCRR资助的中心拨款提供。次级项目的主要支助 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 为子项目列出的总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 肌萎缩侧索硬化症（ALS）是一种使人衰弱的神经退行性疾病，其特征在于 失去运动神经元导致瘫痪并最终死亡家族性肌萎缩侧索硬化症是由 含铜和锌的酶超氧化物歧化酶（SOD 1）突变，这是一种重要的 抗氧化蛋白患者的脊髓含有丰富的富含蛋白质的聚集体， 这表明，与其他神经退行性疾病一样，SOD 1相关的ALS是一种蛋白质错误折叠， 疾病最近的研究表明，未修饰的全长SOD 1是主要的 这些聚集体中的蛋白质，其外观与症状的发作相关。是 目前尚不清楚聚集体中的SOD 1蛋白是否保留了其Cu/Zn金属离子 状态或金属离子的丢失是否导致蛋白质错误折叠。由于SOD 1依赖于Cu 而锌因其活性和稳定性，是疾病检查的重要成分。 在这项研究中，我们将检查ALS小鼠的脊髓，这些小鼠产生一系列SOD 1， 一些突变在体外结合金属，另一些则不结合。转基因 过表达突变形式的SOD 1的小鼠在脊髓中形成聚集体， 病理学包括瘫痪。利用X射线荧光微探针，我们将测定（1）Cu/Zn 聚集体内的Cu含量，（2）聚集体内的Cu氧化态，和（3）聚集体内的Cu氧化态。 未受影响神经元的Cu/Zn含量。这些数据将与FTIR叠加并关联 提供聚集β折叠分布的显微镜图像（在NSLS收集） 蛋白质和脂质过氧化。确定蛋白质代谢状态， 聚集体形成和神经元细胞死亡对于理解ALS病理学至关重要， 随后可能导致开发治疗或治愈ALS的药物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Amyotrophic lateral sclerosis (ALS) is a debilitating neurodegenerative disease characterized by loss of motor neurons leading to paralysis and ultimately death. Familial ALS is caused by mutations in the copper- and zinc-containing enzyme superoxide dismutase (SOD1), a vital antioxidant protein. The spinal cords of patients contain abundant protein-rich aggregates, suggesting that SOD1-linked ALS, like other neurodegenerative diseases, is a proteinmisfolding disease. Recently it was shown that unmodified, full-length SOD1 is the principal protein in these aggregates, whose appearance correlates with the onset of symptoms. It is currently not known whether the SOD1 protein in the aggregates retains its Cu/Zn metallation state or whether loss of the metal ions results in protein misfolding. As SOD1 relies on Cu and Zn for its activity and stability, it is an important component of the disease to examine. In this study, we will examine the spinal cords of ALS mice that produce a range of SOD1 mutations ¿ some mutations that bind metal in vitro and others that do not. The transgenic mice that overexpress mutant forms of SOD1 develop aggregates in the spinal cord and disease pathology including paralysis. Using the XRF microprobe, we will determine (1) the Cu/Zn content within the aggregates, (2) the Cu oxidation state within the aggregates, and (3) the Cu/Zn content in unaffected neurons. These data will be superimposed and correlated with FTIR microscopy images (collected at the NSLS) that provide a distribution of aggregated beta sheet protein and lipid peroxidation. Determining the link between protein metallation state, aggregate formation, and neuron cell death is critical for understanding ALS pathology and could subsequently lead to developing a drug to treat or cure ALS.