Single particle detection and characterization of synuclein co-aggregation
Single particle detection and characterization of synuclein co-aggregation
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DOI：
10.1016/j.bbrc.2005.06.025
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发表时间：
2005-08-12
影响因子：
3.1
通讯作者：
Kretzschmar, H
中科院分区：
文献类型：
--
作者：
Giese, A;Bader, B;Kretzschmar, H
蛋白质聚集是许多人类疾病如阿尔茨海默病和帕金森病的关键事件。我们提出了一种通用的方法来定量和表征蛋白质聚集体的双色扫描强荧光靶（SIFT）。除了高灵敏度，这种方法提供了一个独特的机会，研究共聚集过程。由于可以在均相测定中分别分析每种聚集体的两种荧光标记组分的比例，因此即使在含有不同类型聚集体混合物的样品中也可以研究聚集体的分子组成。使用这种方法，我们可以证明野生型α-突触核蛋白与家族性帕金森病中发现的突变变体形成共聚集体。此外，我们发现在非等摩尔混合比下聚集体形成显著增加，这可能具有重要的治疗意义，因为降低异常蛋白质的相对量可能导致蛋白质聚集增加，从而导致不良反应。(c)2005年爱思唯尔公司All rights reserved.
Protein aggregation is the key event in a number of human diseases such as Alzheimer's and Parkinson's disease. We present a general method to quantify and characterize protein aggregates by dual-colour scanning for intensely fluorescent targets (SIFT). In addition to high sensitivity, this approach offers a unique opportunity to study co-aggregation processes. As the ratio of two fluorescently labelled components can be analysed for each aggregate separately in a homogeneous assay, the molecular composition of aggregates can be studied even in samples containing a mixture of different types of aggregates. Using this method, we could show that wild-type alpha-synuclein forms co-aggregates with a mutant variant found in familial Parkinson's disease. Moreover, we found a striking increase in aggregate formation at non-equimolar mixing ratios, which may have important therapeutic implications, as lowering the relative amount of aberrant protein may cause an increase of protein aggregation leading to adverse effects. (c) 2005 Elsevier Inc. All rights reserved.