Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i>
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 intracell...
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MDPI AG
2022-06-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/11/7/1292 |
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| author | Qiaoli Yu Mingxue Ran Yuqing Yang Huaiwei Liu Luying Xun Yongzhen Xia |
| author_facet | Qiaoli Yu Mingxue Ran Yuqing Yang Huaiwei Liu Luying Xun Yongzhen Xia |
| author_sort | Qiaoli Yu |
| collection | DOAJ |
| description | 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 <i>Escherichia coli</i>. 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 <i>tris</i>(2-carboxyethyl)phosphine did not. Among different sulfur-bearing compounds, NaHS, cysteine, S<sub>2</sub>O<sub>3</sub><sup>2−</sup> and diallyl disulfide increased iSS, of which only S<sub>2</sub>O<sub>3</sub><sup>2−</sup> did not inhibit <i>E. coli</i> growth at 10 mM or less. Thus, with the improved method, we have identified reagents that may be used to change iSS in <i>E. coli</i> and other organisms, providing tools to further study the physiological functions of iSS. |
| first_indexed | 2024-03-09T12:19:26Z |
| format | Article |
| id | doaj.art-00760e229ad244b0abafa643fefd01e6 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-09T12:19:26Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-00760e229ad244b0abafa643fefd01e62023-11-30T22:42:14ZengMDPI AGAntioxidants2076-39212022-06-01117129210.3390/antiox11071292Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i>Qiaoli Yu0Mingxue Ran1Yuqing Yang2Huaiwei Liu3Luying Xun4Yongzhen Xia5State Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao 266237, ChinaSulfane 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 <i>Escherichia coli</i>. 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 <i>tris</i>(2-carboxyethyl)phosphine did not. Among different sulfur-bearing compounds, NaHS, cysteine, S<sub>2</sub>O<sub>3</sub><sup>2−</sup> and diallyl disulfide increased iSS, of which only S<sub>2</sub>O<sub>3</sub><sup>2−</sup> did not inhibit <i>E. coli</i> growth at 10 mM or less. Thus, with the improved method, we have identified reagents that may be used to change iSS in <i>E. coli</i> and other organisms, providing tools to further study the physiological functions of iSS.https://www.mdpi.com/2076-3921/11/7/1292intracellular sulfane sulfurthiosulfateredox homeostasissulfur-bearing compounds<i>Escherichia coli</i> |
| spellingShingle | Qiaoli Yu Mingxue Ran Yuqing Yang Huaiwei Liu Luying Xun Yongzhen Xia Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i> Antioxidants intracellular sulfane sulfur thiosulfate redox homeostasis sulfur-bearing compounds <i>Escherichia coli</i> |
| title | Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i> |
| title_full | Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i> |
| title_fullStr | Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i> |
| title_full_unstemmed | Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i> |
| title_short | Optimization of a Method for Detecting Intracellular Sulfane Sulfur Levels and Evaluation of Reagents That Affect the Levels in <i>Escherichia coli</i> |
| title_sort | optimization of a method for detecting intracellular sulfane sulfur levels and evaluation of reagents that affect the levels in i escherichia coli i |
| topic | intracellular sulfane sulfur thiosulfate redox homeostasis sulfur-bearing compounds <i>Escherichia coli</i> |
| url | https://www.mdpi.com/2076-3921/11/7/1292 |
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