Vollers Sabrina S, Teplow David B, Bitan Gal
Center for Neurologic Diseases, Brigham and Women's Hospital and Department of Neurology, Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2005;299:11-8. doi: 10.1385/1-59259-874-9:011.
The assembly of the amyloid beta-protein (Abeta) into neurotoxic oligomers and fibrils is a seminal pathogenic process in Alzheimer's disease (AD). Understanding the mechanisms of Abeta assembly could prove useful in the identification of therapeutic targets. Owing to the metastable nature of Abeta oligomers, it is difficult to obtain interpretable data through application of classical methods, such as electrophoresis, chromatography, fluorescence, and light scattering. Here, we apply the method Photo-Induced Crosslinking of Unmodified Proteins (PICUP) to the study of Abeta oligomerization. This method directly produces covalent bonds among unmodified polypeptide chains through in situ generation of peptide free radicals. PICUP provides a snapshot of the native oligomerization state of proteins and can be used for assembly state analysis of a wide variety of peptides and proteins.
淀粉样β蛋白（Aβ）组装成神经毒性寡聚体和原纤维是阿尔茨海默病（AD）中的一个关键致病过程。了解Aβ组装机制可能有助于确定治疗靶点。由于Aβ寡聚体的亚稳态性质，通过应用经典方法，如电泳、色谱、荧光和光散射，很难获得可解释的数据。在这里，我们将未修饰蛋白质的光诱导交联（PICUP）方法应用于Aβ寡聚化研究。该方法通过原位产生肽自由基直接在未修饰的多肽链之间形成共价键。PICUP提供了蛋白质天然寡聚化状态的快照，可用于多种肽和蛋白质的组装状态分析。
