Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in Escherichia coli.
Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in Escherichia coli.
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大肠杆菌细胞内硫烷硫水平检测方法的优化及影响水平的试剂评价
DOI：
10.3390/antiox11071292
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发表时间：
2022-06-29
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
硫烷硫是一类含有零价硫的化合物。大多数硫烷硫化合物具有反应性并发挥重要的信号作用。涉及硫烷硫的产生和代谢的关键酶已得到表征；然而，人们对如何改变细胞内硫烷硫 (iSS) 水平知之甚少。为了准确测量 iSS，我们优化了之前报道的方法，其中反应性 iSS 与亚硫酸盐反应生成硫代硫酸盐（一种稳定的硫烷硫化合物），然后进行检测。利用改进的方法，测试了影响大肠杆菌 iSS 的几个因素。温度、pH 值和渗透压几乎没有影响。在常用浓度下，大多数测试的氧化剂，包括过氧化氢、叔丁基过氧化氢、次氯酸和二酰胺，不会影响 iSS，但羰基氰化物间氯苯腙会增加 iSS。对于还原剂，10 mM 二硫苏糖醇显着降低 iSS，但三(2-羧乙基)膦则没有。在不同的含硫化合物中，NaHS、半胱氨酸、S2O32−和二烯丙基二硫化物可增加iSS，其中只有S2O32−在10 mM或更低浓度时不能抑制大肠杆菌生长。因此，通过改进的方法，我们已经确定了可用于改变大肠杆菌和其他生物体中iSS的试剂，为进一步研究iSS的生理功能提供了工具。
Sulfane sulfur is a class of compounds containing zero-valent sulfur. Most sulfane sulfur compounds are reactive and play important signaling roles. Key enzymes involved in the production and metabolism of sulfane sulfur have been characterized; however, little is known about how to change intracellular sulfane sulfur (iSS) levels. To accurately measure iSS, we optimized a previously reported method, in which reactive iSS reacts with sulfite to produce thiosulfate, a stable sulfane sulfur compound, before detection. With the improved method, several factors were tested to influence iSS in Escherichia coli. Temperature, pH, and osmotic pressure showed little effect. At commonly used concentrations, most tested oxidants, including hydrogen peroxide, tert-butyl hydroperoxide, hypochlorous acid, and diamide, did not affect iSS, but carbonyl cyanide m-chlorophenyl hydrazone increased iSS. For reductants, 10 mM dithiothreitol significantly decreased iSS, but tris(2-carboxyethyl)phosphine did not. Among different sulfur-bearing compounds, NaHS, cysteine, S2O32− and diallyl disulfide increased iSS, of which only S2O32− did not inhibit E. coli growth at 10 mM or less. Thus, with the improved method, we have identified reagents that may be used to change iSS in E. coli and other organisms, providing tools to further study the physiological functions of iSS.
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影响因子：
6
作者：
Gregory GJ;Boyd EF
通讯作者：
Boyd EF
DOI：
10.2174/1874285800802010018
发表时间：
2008
期刊：
The open microbiology journal
影响因子：
--
作者：
Cheng H;Donahue JL;Battle SE;Ray WK;Larson TJ
通讯作者：
Larson TJ
影响因子：
5.1
作者：
Corona, Fernando;Luis Martinez, Jose;Nikel, Pablo, I
通讯作者：
Nikel, Pablo, I
影响因子：
4.3
作者：
Bruska, Marta K.;Stiebritz, Martin T.;Reiher, Markus
通讯作者：
Reiher, Markus
影响因子：
3.8
作者：
Chiang, Yi-Hsuan;Jen, Lin-Ni;Liu, Cheng-Tzu
通讯作者：
Liu, Cheng-Tzu