Abstract:
Bacteria in nature and as pathogens commonly face oxidative stress which causes damage to proteins, lipids and DNA. This damage is produced by the action of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2), singlet oxygen, superoxide anion and hydroxyl radical. ROS are generated by antimicrobials, environmental factors (e.g., ultraviolet radiation, osmotic stress), aerobic respiration, and host phagocytes during infective processes. Pseudomonas aeruginosa, a versatile bacterium, is a prevalent opportunistic human pathogen which possesses several defense strategies against ROS. Among them, two catalases (KatA and KatB) have been well characterized by their role on the defense against multiple types of stress. In this protocol, KatA and KatB activities are detected by polyacrylamide gel electrophoresis (PAGE). It is also suggested that the detection of KatB is elusive.
摘要:
自然界中的细菌和作为病原体的细菌通常面临氧化应激,这会导致蛋白质损伤,脂质和DNA。这种损害是由活性氧(ROS)如过氧化氢(H2O2)的作用产生的,单线态氧,超氧阴离子和羟基自由基。ROS是由抗菌药物产生的,环境因素(例如,紫外线辐射,渗透应力),有氧呼吸,和感染过程中的宿主吞噬细胞。铜绿假单胞菌,一种多才多艺的细菌,是一种流行的机会性人类病原体,具有多种针对ROS的防御策略。其中,两种过氧化氢酶(KatA和KatB)的特征在于它们在防御多种类型的压力方面的作用。在这个协议中,通过聚丙烯酰胺凝胶电泳(PAGE)检测KatA和KatB活性。还表明KatB的检测是难以捉摸的。