通过聚丙烯酰胺凝胶电泳（PAGE）检测来自……的细胞提取物中的过氧化氢酶活性
Detection of Catalase Activity by Polyacrylamide Gel Electrophoresis (PAGE)in Cell Extracts from .
作者信息
Pezzoni Magdalena, Pizarro Ramón A, Costa Cristina S
机构信息
Dpto. de Radiobiología, Comisión Nacional de Energía Atómica, General San Martín, Argentina.
出版信息
Bio Protoc. 2018 Jun 5;8(11):e2869. doi: 10.21769/BioProtoc.2869.
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 (HO), singlet oxygen, superoxide anion and hydroxyl radical. ROS are generated by antimicrobials, environmental factors (, ultraviolet radiation, osmotic stress), aerobic respiration, and host phagocytes during infective processes. , 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.
自然界中的细菌以及作为病原体的细菌通常会面临氧化应激，氧化应激会对蛋白质、脂质和DNA造成损害。这种损害是由活性氧（ROS）如过氧化氢（HO）、单线态氧、超氧阴离子和羟基自由基的作用产生的。ROS由抗菌剂、环境因素（如紫外线辐射、渗透应激）、有氧呼吸以及感染过程中的宿主吞噬细胞产生。大肠杆菌，一种多能细菌，是一种普遍的机会性人类病原体，它拥有多种抵御ROS的防御策略。其中，两种过氧化氢酶（KatA和KatB）因其在抵御多种类型应激中的作用而得到了充分的表征。在本实验方案中，通过聚丙烯酰胺凝胶电泳（PAGE）检测KatA和KatB的活性。还表明KatB的检测很困难。
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