Kim Heejung, Seong Jihye
Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul 02792, Korea.
Department of Converging Science and Technology, Kyung Hee University, Seoul 02453, Korea.
Materials (Basel). 2021 Jun 2;14(11):3019. doi: 10.3390/ma14113019.
Autophagy is an essential cellular process of self-degradation for dysfunctional or unnecessary cytosolic constituents and organelles. Dysregulation of autophagy is thus involved in various diseases such as neurodegenerative diseases. To investigate the complex process of autophagy, various biochemical, chemical assays, and imaging methods have been developed. Here we introduce various methods to study autophagy, in particular focusing on the review of designs, principles, and limitations of the fluorescent protein (FP)-based autophagy biosensors. Different physicochemical properties of FPs, such as pH-sensitivity, stability, brightness, spectral profile, and fluorescence resonance energy transfer (FRET), are considered to design autophagy biosensors. These FP-based biosensors allow for sensitive detection and real-time monitoring of autophagy progression in live cells with high spatiotemporal resolution. We also discuss future directions utilizing an optobiochemical strategy to investigate the in-depth mechanisms of autophagy. These cutting-edge technologies will further help us to develop the treatment strategies of autophagy-related diseases.
自噬是一种重要的细胞自我降解过程，用于处理功能失调或不必要的胞质成分和细胞器。因此，自噬失调与多种疾病有关，如神经退行性疾病。为了研究自噬的复杂过程，人们开发了各种生化、化学检测方法和成像方法。在这里，我们介绍各种研究自噬的方法，特别侧重于对基于荧光蛋白（FP）的自噬生物传感器的设计、原理和局限性的综述。在设计自噬生物传感器时，会考虑荧光蛋白的不同物理化学性质，如pH敏感性、稳定性、亮度、光谱特征和荧光共振能量转移（FRET）。这些基于荧光蛋白的生物传感器能够以高时空分辨率在活细胞中灵敏地检测和实时监测自噬进程。我们还讨论了利用光生物化学策略深入研究自噬机制的未来方向。这些前沿技术将进一步帮助我们制定自噬相关疾病的治疗策略。
