A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria
The Trx and Grx systems, two disulfide reductase systems, play critical roles in various cell activities. There are great differences between the thiol redox systems in prokaryotes and mammals. Though fluorescent probes have been widely used to detect these systems in mammalian cells. Very few metho...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/11/2/377 |
| _version_ | 1797483220810334208 |
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| author | Ying Zhao Xin Zuo Shuang Liu Wenjun Qian Xuewen Tang Jun Lu |
| author_facet | Ying Zhao Xin Zuo Shuang Liu Wenjun Qian Xuewen Tang Jun Lu |
| author_sort | Ying Zhao |
| collection | DOAJ |
| description | The Trx and Grx systems, two disulfide reductase systems, play critical roles in various cell activities. There are great differences between the thiol redox systems in prokaryotes and mammals. Though fluorescent probes have been widely used to detect these systems in mammalian cells. Very few methods are available to detect rapid changes in the redox systems of prokaryotes. Here we investigated whether Fast-TRFS, a disulfide-containing fluorescent probe utilized in analysis of mammalian thioredoxin reductase, could be used to detect cellular disulfide reducibility in bacteria. Fast-TRFS exhibited good substrate qualities for both bacterial thioredoxin and GSH-glutaredoxin systems in vitro, with Trx system having higher reaction rate. Moreover, the Fast-TRFS was used to detect the disulfide reductase activity in various bacteria and redox-related gene null <i>E. coli</i>. Some glutaredoxin-deficient bacteria had stronger fast disulfide reducibility. The Trx system was shown to be the predominant disulfide reductase for fast disulfide reduction rather than the Grx system. These results demonstrated that Fast-TRFS is a viable probe to detect thiol-dependent disulfide reductases in bacteria. It also indicated that cellular disulfide reduction could be classified into fast and slow reaction, which are predominantly catalyzed by <i>E. coli</i> Trx and Grx system, respectively. |
| first_indexed | 2024-03-09T22:43:49Z |
| format | Article |
| id | doaj.art-bcf6267ef0e54f6fa098701cbaf5b450 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-09T22:43:49Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-bcf6267ef0e54f6fa098701cbaf5b4502023-11-23T18:32:52ZengMDPI AGAntioxidants2076-39212022-02-0111237710.3390/antiox11020377A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in BacteriaYing Zhao0Xin Zuo1Shuang Liu2Wenjun Qian3Xuewen Tang4Jun Lu5Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education (Southwest University), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaKey Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education (Southwest University), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaKey Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education (Southwest University), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaKey Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education (Southwest University), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaKey Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education (Southwest University), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaKey Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education (Southwest University), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaThe Trx and Grx systems, two disulfide reductase systems, play critical roles in various cell activities. There are great differences between the thiol redox systems in prokaryotes and mammals. Though fluorescent probes have been widely used to detect these systems in mammalian cells. Very few methods are available to detect rapid changes in the redox systems of prokaryotes. Here we investigated whether Fast-TRFS, a disulfide-containing fluorescent probe utilized in analysis of mammalian thioredoxin reductase, could be used to detect cellular disulfide reducibility in bacteria. Fast-TRFS exhibited good substrate qualities for both bacterial thioredoxin and GSH-glutaredoxin systems in vitro, with Trx system having higher reaction rate. Moreover, the Fast-TRFS was used to detect the disulfide reductase activity in various bacteria and redox-related gene null <i>E. coli</i>. Some glutaredoxin-deficient bacteria had stronger fast disulfide reducibility. The Trx system was shown to be the predominant disulfide reductase for fast disulfide reduction rather than the Grx system. These results demonstrated that Fast-TRFS is a viable probe to detect thiol-dependent disulfide reductases in bacteria. It also indicated that cellular disulfide reduction could be classified into fast and slow reaction, which are predominantly catalyzed by <i>E. coli</i> Trx and Grx system, respectively.https://www.mdpi.com/2076-3921/11/2/377thioredoxinglutathioneredoxbacteriaprobefast disulfide reduction |
| spellingShingle | Ying Zhao Xin Zuo Shuang Liu Wenjun Qian Xuewen Tang Jun Lu A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria Antioxidants thioredoxin glutathione redox bacteria probe fast disulfide reduction |
| title | A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria |
| title_full | A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria |
| title_fullStr | A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria |
| title_full_unstemmed | A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria |
| title_short | A Fluorescent Probe to Detect Quick Disulfide Reductase Activity in Bacteria |
| title_sort | fluorescent probe to detect quick disulfide reductase activity in bacteria |
| topic | thioredoxin glutathione redox bacteria probe fast disulfide reduction |
| url | https://www.mdpi.com/2076-3921/11/2/377 |
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