Department of Molecular Mechanisms of Disease (DMMD), University of Zurich, Zurich, Switzerland.
Methods Mol Biol. 2021;2276:165-171. doi: 10.1007/978-1-0716-1266-8_12.
ADP-ribosylation is a posttranslational protein modification, involved in various cellular processes, ranging from DNA-damage repair to apoptosis. While its function has been studied amply with respect to genotoxic stress-associated nuclear ADP-ribosylation, the functional relevance of mitochondrial ADP-ribosylation remains so far poorly studied. This is mainly attributed to the absence of powerful techniques able to detect the modification. However, the usage of recently developed anti-ADP-ribose-specific antibodies allows now to investigate mitochondrial ADP-ribosylation under physiological and pathophysiological conditions. In the below method, we describe in detail how to efficiently detect and quantify mitochondrial ADP-ribosylation via immunofluorescence.
ADP-核糖基化是一种翻译后蛋白质修饰，参与多种细胞过程，从 DNA 损伤修复到细胞凋亡。虽然其功能已在与遗传毒性应激相关的核 ADP-核糖基化方面得到了充分研究，但线粒体 ADP-核糖基化的功能相关性至今仍研究甚少。这主要归因于缺乏能够检测修饰的强大技术。然而，最近开发的抗 ADP-核糖基特异性抗体的使用现在允许在生理和病理生理条件下研究线粒体 ADP-核糖基化。在下面的方法中，我们详细描述了如何通过免疫荧光术有效地检测和定量线粒体 ADP-核糖基化。
