From the Department of Molecular and Pharmacological Neuroscience, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima 734-8551.
National Institute of Agrobiological Sciences, Ohwashi 1-2, Tsukuba, Ibaraki 305-8634.
J Biol Chem. 2010 Oct 22;285(43):33252-33264. doi: 10.1074/jbc.M110.146704. Epub 2010 Aug 12.
Several missense mutations in the protein kinase Cγ (γPKC) gene have been found to cause spinocerebellar ataxia type 14 (SCA14), an autosomal dominant neurodegenerative disease. We previously demonstrated that the mutant γPKC found in SCA14 is susceptible to aggregation, which induces apoptotic cell death. The disaccharide trehalose has been reported to inhibit aggregate formation and to alleviate symptoms in cellular and animal models of Huntington disease, Alzheimer disease, and prion disease. Here, we show that trehalose can be incorporated into SH-SY5Y cells and reduces the aggregation of mutant γPKC-GFP, thereby inhibiting apoptotic cell death in SH-SY5Y cells and primary cultured Purkinje cells (PCs). Trehalose acts by directly stabilizing the conformation of mutant γPKC without affecting protein turnover. Trehalose was also found to alleviate the improper development of dendrites in PCs expressing mutant γPKC-GFP without aggregates but not in PCs with aggregates. In PCs without aggregates, trehalose improves the mobility and translocation of mutant γPKC-GFP, probably by inhibiting oligomerization and thereby alleviating the improper development of dendrites. These results suggest that trehalose counteracts various cellular dysfunctions that are triggered by mutant γPKC in both neuronal cell lines and primary cultured PCs by inhibiting oligomerization and aggregation of mutant γPKC.
几种蛋白激酶 Cγ(γPKC)基因突变已被发现可导致脊髓小脑共济失调 14 型(SCA14),这是一种常染色体显性神经退行性疾病。我们之前证明,在 SCA14 中发现的突变 γPKC 容易聚集,从而诱导细胞凋亡。二糖海藻糖已被报道可抑制聚集物的形成,并在亨廷顿病、阿尔茨海默病和朊病毒病的细胞和动物模型中减轻症状。在这里,我们表明海藻糖可以被整合到 SH-SY5Y 细胞中,并减少突变体 γPKC-GFP 的聚集,从而抑制 SH-SY5Y 细胞和原代培养的浦肯野细胞(PCs)中的细胞凋亡。海藻糖通过直接稳定突变体 γPKC 的构象起作用,而不影响蛋白质周转。还发现海藻糖可减轻表达突变体 γPKC-GFP 的 PCs 中无聚集体的树突异常发育,但对有聚集体的 PCs 无作用。在没有聚集体的 PCs 中,海藻糖可改善突变体 γPKC-GFP 的流动性和易位,可能通过抑制寡聚化从而减轻树突的异常发育。这些结果表明,海藻糖通过抑制突变体 γPKC 的寡聚化和聚集,在神经元细胞系和原代培养的 PCs 中对抗由突变体 γPKC 触发的各种细胞功能障碍。