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Human serum albumin-derived biosensor for visualizing copper(II) and hydrogen sulfide interactions in cancer cells
Yong An1, Yucen Zhou2, Linghua Zhang3
1The First School of Clinical Medical, Gansu University of Chinese Medicine, Lanzhou, Gansu 730000, PR China.
Introduction:
Accurate monitoring of Cu2+ and H2S levels is one of the most reliable methods for diagnosing tumors and assessing treatment efficacy. However, the development of fluorescent probes capable of detecting Cu2+ and H2S in cancer cells against interference from the tumour microenvironment remains a challenge.
Objective:
This study aimed to develop a novel peptide-based biosensor for monitoring Cu2+ and H2S in cancer cells and visualizing the effects of Cu2+-H2S interactions on redox homeostasis.
Methods:
First, we designed and synthesized the sensor NBD-DAH (NBD-Asp-Ala-His) by mimicking the Cu2+ binding site of human serum albumin (HSA). Second, we evaluated the specificity, sensitivity, and resistance of NBD-DAH to interference from the biological environment in the sequential detection of Cu2+ and H2S. Third, we investigated the binding sites and sensing mechanism between NBD-DAH and Cu2+. Finally, we evaluated the potential and biosafety of NBD-DAH for monitoring Cu2+ and H2S, visualizing their interactions in cancer cells, and exploring the effects and potential mechanisms of these interactions on redox homeostasis.
Results:
NBD-DAH can specifically and sensitively detect Cu2+ and H2S in sequence through an "on-off-on" fluorescence response. Because of its high cell membrane permeability, broad pH compatibility, and rapid response, the sensor resists interferences from the complex tumor microenvironment and enables reliable detection of Cu2+ and H2S level changes in cancer cells. Furthermore, live-cell imaging with NBD-DAH revealed a molecular mechanism in which H2S specifically binds to Cu2+ in cancer cells, leading to the depletion of superoxide dismutase 1 (SOD1) activity and subsequent regulation of redox homeostasis.
Conclusion:
We developed a novel biosensor, NBD-DAH, and successfully used it to monitor Cu2+ and H2S levels and visualize their interactions in cancer cells. The interaction between Cu2+ and H2S influenced intracellular redox homeostasis, potentially through its effect on SOD1 activity.
