A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur
Reactive sulfane sulfur, including persulfide and polysulfide, is a type of regular cellular component, playing an antioxidant role. Its function may be organelle-dependent; however, the shortage of probes for detecting organellar reactive sulfane sulfur has hindered further investigation. Herein, w...
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MDPI AG
2020-10-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/9/10/985 |
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| author | Zimai Li Qingda Wang Yongzhen Xia Luying Xun Huaiwei Liu |
| author_facet | Zimai Li Qingda Wang Yongzhen Xia Luying Xun Huaiwei Liu |
| author_sort | Zimai Li |
| collection | DOAJ |
| description | Reactive sulfane sulfur, including persulfide and polysulfide, is a type of regular cellular component, playing an antioxidant role. Its function may be organelle-dependent; however, the shortage of probes for detecting organellar reactive sulfane sulfur has hindered further investigation. Herein, we reported a red fluorescent protein (mCherry)-based probe for specifically detecting intracellular reactive sulfane sulfur. By mutating two amino acid residues of mCherry (A150 and S151) to cysteine residues, we constructed a mCherry mutant, which reacted with reactive sulfane sulfur to form an intramolecular –S<sub>n</sub>– bond (<i>n</i> ≥ 3). The bond largely decreased the intensity of 610 nm emission (excitation at 587 nm) and slightly increased the intensity of 466 nm emission (excitation at 406 nm). The 466/610 nm emission ratio was used to indicate the relative abundance of reactive sulfane sulfur. We then expressed this mutant in the cytoplasm and mitochondria of <i>Saccharomyces cerevisiae</i>. The 466/610 nm emission ratio revealed that mitochondria had a higher level of reactive sulfane sulfur than cytoplasm. Thus, the mCherry mutant can be used as a specific probe for detecting reactive sulfane sulfur in vivo. |
| first_indexed | 2024-03-10T15:39:12Z |
| format | Article |
| id | doaj.art-1daf3a1e8b774a2fbd5c2e20ac76cc01 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-10T15:39:12Z |
| publishDate | 2020-10-01 |
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| series | Antioxidants |
| spelling | doaj.art-1daf3a1e8b774a2fbd5c2e20ac76cc012023-11-20T16:57:14ZengMDPI AGAntioxidants2076-39212020-10-0191098510.3390/antiox9100985A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane SulfurZimai Li0Qingda Wang1Yongzhen Xia2Luying Xun3Huaiwei Liu4State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, ChinaReactive sulfane sulfur, including persulfide and polysulfide, is a type of regular cellular component, playing an antioxidant role. Its function may be organelle-dependent; however, the shortage of probes for detecting organellar reactive sulfane sulfur has hindered further investigation. Herein, we reported a red fluorescent protein (mCherry)-based probe for specifically detecting intracellular reactive sulfane sulfur. By mutating two amino acid residues of mCherry (A150 and S151) to cysteine residues, we constructed a mCherry mutant, which reacted with reactive sulfane sulfur to form an intramolecular –S<sub>n</sub>– bond (<i>n</i> ≥ 3). The bond largely decreased the intensity of 610 nm emission (excitation at 587 nm) and slightly increased the intensity of 466 nm emission (excitation at 406 nm). The 466/610 nm emission ratio was used to indicate the relative abundance of reactive sulfane sulfur. We then expressed this mutant in the cytoplasm and mitochondria of <i>Saccharomyces cerevisiae</i>. The 466/610 nm emission ratio revealed that mitochondria had a higher level of reactive sulfane sulfur than cytoplasm. Thus, the mCherry mutant can be used as a specific probe for detecting reactive sulfane sulfur in vivo.https://www.mdpi.com/2076-3921/9/10/985sulfane sulfurantioxidationmCherrymitochondria<i>Saccharomyces cerevisiae</i> |
| spellingShingle | Zimai Li Qingda Wang Yongzhen Xia Luying Xun Huaiwei Liu A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur Antioxidants sulfane sulfur antioxidation mCherry mitochondria <i>Saccharomyces cerevisiae</i> |
| title | A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur |
| title_full | A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur |
| title_fullStr | A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur |
| title_full_unstemmed | A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur |
| title_short | A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur |
| title_sort | red fluorescent protein based probe for detection of intracellular reactive sulfane sulfur |
| topic | sulfane sulfur antioxidation mCherry mitochondria <i>Saccharomyces cerevisiae</i> |
| url | https://www.mdpi.com/2076-3921/9/10/985 |
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