Scherz-Shouval Ruth, Elazar Zvulun
Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel.
Methods Enzymol. 2009;452:119-30. doi: 10.1016/S0076-6879(08)03608-2.
Reactive oxygen species (ROS) are potentially harmful to cells because of their ability to oxidize cell constituents such as DNA, proteins, and lipids. However, at low levels, and under tight control, this feature makes them excellent modifiers in a variety of signal transduction pathways, including autophagy. Autophagy was traditionally associated with oxidative stress, acting in the degradation of oxidized proteins and organelles. Recently, a signaling role was suggested for ROS in the regulation of autophagy, leading, under different circumstances, either to survival or to death. To study the effects of ROS on this pathway, one must determine the localization, intensity, kinetics, and essentiality of the oxidative signal in autophagy. Moreover, once characterized, detection and manipulation of ROS formation could be used to monitor and control autophagic activity. In this chapter we discuss methods to examine ROS in the context of autophagy.
活性氧（ROS）因其能够氧化细胞成分（如DNA、蛋白质和脂质）而对细胞具有潜在危害。然而，在低水平且受到严格控制的情况下，这一特性使它们成为包括自噬在内的多种信号转导途径中的优秀调节剂。传统上，自噬与氧化应激相关，作用于氧化蛋白质和细胞器的降解。最近，有人提出ROS在自噬调节中具有信号传导作用，在不同情况下，可导致细胞存活或死亡。为了研究ROS对这一途径的影响，必须确定自噬中氧化信号的定位、强度、动力学和必要性。此外，一旦明确其特征，ROS形成的检测和操纵可用于监测和控制自噬活性。在本章中，我们将讨论在自噬背景下检测ROS的方法。
