A Red Fluorescent Protein-Based Probe for Detection of Intracellular Reactive Sulfane Sulfur

• Main Authors: Zimai Li (Author), Qingda Wang (Author), Yongzhen Xia (Author), Luying Xun (Author), Huaiwei Liu (Author)
• Format: Bog
• Udgivet: MDPI AG, 2020-10-01T00:00:00Z.
• Fag: article
• Online adgang: Connect to this object online.

Summary

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_n- 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.

Emne beskrivelse

10.3390/antiox9100985
2076-3921