Holt Katherine B, Bard Allen J
Department of Chemistry and Biochemistry, University of Texas at Austin, 1 University Station A5300, Austin, Texas 78712-0165, USA.
Biochemistry. 2005 Oct 4;44(39):13214-23. doi: 10.1021/bi0508542.
Electrochemical techniques were used to study the behavior of Escherichia coli on the addition of <or=10 microM AgNO(3). Respiration in the presence of glucose was measured using a Clark ultramicroelectrode to determine the oxygen concentration as a function of time. The rate of respiration increased initially upon the addition of silver(I) because of the uncoupling of the respiratory chain, followed by cessation of respiration. The toxicity of 1 microM AgNO(3), as determined by the time until respiration ceased, increased in the absence of glucose and in the presence of K(+). The uptake of 1 microM Ag(+), measured from the stripping peak height of Ag electrodeposited on a 25 microm Pt ultramicroelectrode, showed a fastest uptake in the presence of glucose, although glucose was not required for uptake. Efflux of Ag(+) from cells was also detected in the presence of glucose. Scanning electrochemical microscopy (SECM) was used to follow the uptake of 1 microM Ag(+) by living and dead E. coli immobilized on a glass slide coated with poly-l-lysine. Ferricyanide was used as an alternative electron acceptor to oxygen to probe the site of inhibition of Ag(+) in the respiratory chain.
采用电化学技术研究了添加≤10微摩尔硝酸银时大肠杆菌的行为。使用克拉克超微电极测量葡萄糖存在下的呼吸作用,以确定氧浓度随时间的变化。添加银离子(I)后,由于呼吸链解偶联,呼吸速率最初增加,随后呼吸停止。由呼吸停止时间确定的1微摩尔硝酸银的毒性,在无葡萄糖和有钾离子存在时增加。从沉积在25微米铂超微电极上的银的溶出峰高测量1微摩尔银离子(+)的摄取量,结果表明在有葡萄糖存在时摄取最快,尽管摄取并不需要葡萄糖。在有葡萄糖存在时也检测到银离子(+)从细胞中流出。使用扫描电化学显微镜(SECM)跟踪固定在涂有聚-L-赖氨酸的载玻片上的活的和死的大肠杆菌对1微摩尔银离子(+)的摄取。铁氰化物用作氧的替代电子受体,以探测银离子(+)在呼吸链中的抑制位点。