Organelle Biology and Cellular Ageing Lab, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, Assam, India.
Mol Biol Rep. 2023 Jul;50(7):5547-5556. doi: 10.1007/s11033-023-08457-7. Epub 2023 May 8.
α-synuclein aggregation is the hallmark feature of Parkinson's disease. Both familial and sporadic forms of the disease exhibit this feature. Several mutations have been identified in patients and are associated with the disease pathology.
We have used site-directed mutagenesis to generate α-synuclein mutant variants tagged with GFP. Fluorescence microscopy, flow cytometry, western blotting, cell viability and oxidative stress analysis were performed to investigate the effect of two less studied α-synuclein variants. In this study we characterized two less studied α-synuclein mutations, A18T and A29S, in the well-established yeast model. Our data shows variable expression, distribution and toxicity of the protein in the mutant variants A18T, A29S, A53T and WT. The cells expressing the double mutant variant A18T/A53T showed the most increase in the aggregation phenotype and also depicted reduced viability suggesting a more substantial effect of this variant.
The outcome of our study highlights the variable localization, aggregation phenotype and toxicity of the studied α-synuclein variants. This underscores the importance of in-depth analysis of every disease-associated mutation which may result in variable cellular phenotype.
α-突触核蛋白聚集是帕金森病的标志性特征。该病既有家族性也有散发性形式，都表现出这一特征。已在患者中发现了几种突变，并与疾病病理相关。
我们使用定点诱变生成了带有 GFP 标记的 α-突触核蛋白突变变体。通过荧光显微镜、流式细胞术、Western blot、细胞活力和氧化应激分析来研究两种研究较少的 α-突触核蛋白变体的影响。在这项研究中，我们在成熟的酵母模型中对两种研究较少的 α-突触核蛋白突变 A18T 和 A29S 进行了表征。我们的数据显示，在突变变体 A18T、A29S、A53T 和 WT 中，该蛋白的表达、分布和毒性存在差异。表达双突变变体 A18T/A53T 的细胞表现出聚集表型的显著增加，同时也显示出活力降低，表明该变体的影响更为显著。
我们研究的结果强调了所研究的 α-突触核蛋白变体的可变定位、聚集表型和毒性。这突显了对每个与疾病相关的突变进行深入分析的重要性，因为这可能导致不同的细胞表型。
