Dimerization, Oligomerization, and Aggregation of Human Amyotrophic Lateral Sclerosis Copper/Zinc Superoxide Dismutase 1 Protein Mutant Forms in Live Cells
Dimerization, Oligomerization, and Aggregation of Human Amyotrophic Lateral Sclerosis Copper/Zinc Superoxide Dismutase 1 Protein Mutant Forms in Live Cells
复制标题
DOI：
10.1074/jbc.m113.542613
复制
发表时间：
2014-05-23
影响因子：
4.8
通讯作者：
Grailhe, Regis
中科院分区：
文献类型：
--
作者：
Kim, Jiho;Lee, Honggun;Grailhe, Regis
超过100个铜/锌超氧化物歧化酶1（SOD 1）基因突变已被表征。这些突变导致ALS中运动神经元的死亡。在其天然形式中，SOD 1蛋白在胞质溶胶中表达为同源二聚体。体外研究表明，SOD 1突变损害了蛋白质的二聚动力学，体内研究表明，SOD 1在家族性ALS患者中形成聚集体。在这项研究中，我们通过非侵入性荧光技术分析了WT SOD 1和9种突变（mt）形式的蛋白质。使用显微技术，如荧光共振能量转移，荧光互补，基于图像的定量和荧光相关光谱，我们研究了SOD 1二聚，寡聚和聚集。我们的研究结果表明，SOD 1突变导致SOD 1二聚化的损害，随后，影响蛋白质聚集。我们还表明，SOD 1 WT和mt蛋白可以二聚化。然而，聚集体主要由SOD 1 mt蛋白组成。
More than 100 copper/zinc superoxide dismutase 1 (SOD1) genetic mutations have been characterized. These mutations lead to the death of motor neurons in ALS. In its native form, the SOD1 protein is expressed as a homodimer in the cytosol. In vitro studies have shown that SOD1 mutations impair the dimerization kinetics of the protein, and in vivo studies have shown that SOD1 forms aggregates in patients with familial forms of ALS. In this study, we analyzed WT SOD1 and 9 mutant (mt) forms of the protein by non-invasive fluorescence techniques. Using microscopic techniques such as fluorescence resonance energy transfer, fluorescence complementation, image-based quantification, and fluorescence correlation spectroscopy, we studied SOD1 dimerization, oligomerization, and aggregation. Our results indicate that SOD1 mutations lead to an impairment in SOD1 dimerization and, subsequently, affect protein aggregation. We also show that SOD1 WT and mt proteins can dimerize. However, aggregates are predominantly composed of SOD1 mt proteins.