Laboratorio de Microbiología Molecular, Facultad de Química y Biología, Universidad de Santiago de Chile, Santiago, Chile.
PLoS One. 2007 Feb 14;2(2):e211. doi: 10.1371/journal.pone.0000211.
Biochemical, genetic, enzymatic and molecular approaches were used to demonstrate, for the first time, that tellurite (TeO(3) (2-)) toxicity in E. coli involves superoxide formation. This radical is derived, at least in part, from enzymatic TeO(3) (2-) reduction. This conclusion is supported by the following observations made in K(2)TeO(3)-treated E. coli BW25113: i) induction of the ibpA gene encoding for the small heat shock protein IbpA, which has been associated with resistance to superoxide, ii) increase of cytoplasmic reactive oxygen species (ROS) as determined with ROS-specific probe 2'7'-dichlorodihydrofluorescein diacetate (H(2)DCFDA), iii) increase of carbonyl content in cellular proteins, iv) increase in the generation of thiobarbituric acid-reactive substances (TBARs), v) inactivation of oxidative stress-sensitive [Fe-S] enzymes such as aconitase, vi) increase of superoxide dismutase (SOD) activity, vii) increase of sodA, sodB and soxS mRNA transcription, and viii) generation of superoxide radical during in vitro enzymatic reduction of potassium tellurite.
采用生物化学、遗传学、酶学和分子生物学方法首次证明碲酸盐(TeO3(2-))对大肠杆菌的毒性涉及超氧自由基的形成。这种自由基至少部分来源于酶促的 TeO3(2-)还原。这一结论得到了以下观察结果的支持:在 K2TeO3 处理的大肠杆菌 BW25113 中:i)诱导编码小热休克蛋白 IbpA 的 ibpA 基因的表达,IbpA 与对超氧自由基的抗性有关;ii)用 ROS 特异性探针 2'7'-二氯二氢荧光素二乙酸酯(H2DCFDA)测定细胞质中活性氧物种(ROS)的增加;iii)细胞蛋白中羰基含量的增加;iv)硫代巴比妥酸反应物质(TBARs)的生成增加;v)氧化应激敏感的[Fe-S]酶如 aconitase 的失活;vi)超氧化物歧化酶(SOD)活性的增加;vii)sodA、sodB 和 soxS mRNA 转录的增加;viii)在体外酶促还原碲酸钾时生成超氧自由基。