Department of Chemistry and Biochemistry, University of California, San Diego, CA, United States.
Department of Chemistry and Biochemistry, University of California, San Diego, CA, United States.
Methods Enzymol. 2020;639:91-114. doi: 10.1016/bs.mie.2020.04.010. Epub 2020 Apr 25.
The aggregation of misfolded proteins into amyloids is a common characteristic of many neurodegenerative and non-neurologic diseases. Fluorescent amyloid-targeting probes that discriminate amyloids based on differences in protein composition can provide rapid information to aid in disease diagnosis. In this chapter, we present protocols for the synthesis and use of ANCA-11 as an environmentally-sensitive amyloid-targeting probe that can fluorescently discriminate between amyloids with different disease origin. We also present a protocol for preparing amyloid samples of synthetic Amyloid-β(1-42), as problems with amyloid preparations can be a large driver of time and cost for research. The methods presented here can be generalized for evaluation of other amyloid-targeting fluorescent probes with different aggregates of amyloidogenic proteins in solution or in tissue.
蛋白质错误折叠聚集形成淀粉样纤维是许多神经退行性和非神经疾病的共同特征。基于蛋白质组成差异来区分淀粉样纤维的荧光靶向探针可以提供快速的信息来辅助疾病诊断。在这一章中,我们介绍了 ANCA-11 的合成和使用方案,ANCA-11 是一种对环境敏感的淀粉样纤维靶向探针,可以荧光区分不同来源的淀粉样纤维。我们还介绍了一种用于制备合成的淀粉样β(1-42)(Amyloid-β(1-42))淀粉样纤维样本的方案,因为淀粉样纤维的制备问题可能会极大地增加研究的时间和成本。这里介绍的方法可以推广用于评估溶液或组织中不同淀粉样纤维聚集的其他淀粉样纤维靶向荧光探针。