Detection of Damage of Biomolecules by Free Radicals and Related Reactive Species

Abstract

Lipids, proteins, and nucleic acids are major cellular targets of reactive oxygen and nitrogen species (ROS/RNS). Oxidative damage to these cellular constituents has been increasingly recognized as a significant pathophysiological event leading to many disease processes. Thus, determination of ROS/RNSelicited damage to lipids, proteins, and nuclear acids is crucial for investigating the oxidative mechanisms of human diseases and assessing the efficacy of antioxidant-based therapies. This chapter first introduces the general experimental approaches to assessing the effects of ROS/RNS on cellular biomolecules and then describes the commonly used methods to determine ROS/RNS-mediated biological damage, with an emphasis on the assays that detect lipid peroxidation, protein oxidation, and oxidative DNA base modifications. The chapter ends with a discussion of biomarkers of oxidative stress and their value in assessing disease pathophysiology as well as the efficacy of antioxidant intervention.

Keywords: Biomarkers, DNA base modifications, DNA damage, F2-isoprostanes, Lipid peroxidation, Methodologies, Oxidative stress, Protein oxidation, Reactive nitrogen species, Reactive oxygen species.