Heine Erin M, Berger Tamar R, Pluciennik Anna, Orr Christopher R, Zboray Lori, Merry Diane E
From the Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
From the Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107
J Biol Chem. 2015 May 15;290(20):12572-84. doi: 10.1074/jbc.M114.617894. Epub 2015 Mar 20.
Proteolysis of polyglutamine-expanded proteins is thought to be a required step in the pathogenesis of several neurodegenerative diseases. The accepted view for many polyglutamine proteins is that proteolysis of the mutant protein produces a "toxic fragment" that induces neuronal dysfunction and death in a soluble form; toxicity of the fragment is buffered by its incorporation into amyloid-like inclusions. In contrast to this view, we show that, in the polyglutamine disease spinal and bulbar muscular atrophy, proteolysis of the mutant androgen receptor (AR) is a late event. Immunocytochemical and biochemical analyses revealed that the mutant AR aggregates as a full-length protein, becoming proteolyzed to a smaller fragment through a process requiring the proteasome after it is incorporated into intranuclear inclusions. Moreover, the toxicity-predicting conformational antibody 3B5H10 bound to soluble full-length AR species but not to fragment-containing nuclear inclusions. These data suggest that the AR is toxic as a full-length protein, challenging the notion of polyglutamine protein fragment-associated toxicity by redefining the role of AR proteolysis in spinal and bulbar muscular atrophy pathogenesis.
多聚谷氨酰胺扩展蛋白的蛋白水解被认为是几种神经退行性疾病发病机制中的一个必要步骤。对于许多多聚谷氨酰胺蛋白,公认的观点是突变蛋白的蛋白水解产生一个“毒性片段”,该片段以可溶形式诱导神经元功能障碍和死亡;片段的毒性通过其掺入淀粉样蛋白样包涵体而得到缓冲。与这种观点相反,我们发现,在多聚谷氨酰胺疾病脊髓延髓肌肉萎缩症中,突变雄激素受体(AR)的蛋白水解是一个晚期事件。免疫细胞化学和生化分析表明,突变AR以全长蛋白形式聚集,在其被掺入核内包涵体后,通过一个需要蛋白酶体的过程被蛋白水解为一个较小的片段。此外,预测毒性的构象抗体3B5H10与可溶性全长AR物种结合,但不与含片段的核内包涵体结合。这些数据表明AR作为全长蛋白具有毒性,通过重新定义AR蛋白水解在脊髓延髓肌肉萎缩症发病机制中的作用,挑战了多聚谷氨酰胺蛋白片段相关毒性的概念。