Mathilde Pucher, Kirrthana Makenthirathasan, Hadrien Jalaber, Thomas LeSaux, Oliver Nüsse, Gilles Doisneau, Yann Bourdreux, Blaise Gatin-Fraudet, Ludovic Jullien, Boris Vauzeilles, Dominique Guianvarc'h, Marie Erard, Dominique Urban
{"title":"基于硼酸的荧光探针作为一种替代Amplex Red法实时监测活细胞中H2O2的方法。","authors":"Mathilde Pucher, Kirrthana Makenthirathasan, Hadrien Jalaber, Thomas LeSaux, Oliver Nüsse, Gilles Doisneau, Yann Bourdreux, Blaise Gatin-Fraudet, Ludovic Jullien, Boris Vauzeilles, Dominique Guianvarc'h, Marie Erard, Dominique Urban","doi":"10.1021/acschembio.5c00156","DOIUrl":null,"url":null,"abstract":"<p><p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is a crucial reactive oxygen species (ROS) involved in regulating both physiological and pathological processes. Excessive H<sub>2</sub>O<sub>2</sub> production can lead to oxidative stress, contributing to aging, cancer, and neurodegenerative diseases. In contrast to other ROS exhibiting short lifespans, H<sub>2</sub>O<sub>2</sub> is relatively stable, and its spatial and temporal dynamics are central to understanding its pathophysiological role. Therefore, the development of fluorescent probes that are highly selective, sensitive, and capable of a rapid response is still required. To date, numerous fluorescent probes have been developed. Among them, boronic acid triggers have attracted considerable attention but often suffer from limited reactivity, preventing real-time H<sub>2</sub>O<sub>2</sub> monitoring. To overcome this lack of reactivity, we report the design and synthesis of new borinic acid-based fluorogenic probes for H<sub>2</sub>O<sub>2</sub> detection in cellular environments. These probes are based on a hemicyanine scaffold functionalized with the borinic acid trigger, which demonstrated superior kinetics compared to its boronic counterpart. These probes enable efficient real-time monitoring of H<sub>2</sub>O<sub>2</sub> in cellular models, both extracellularly and intracellularly. The kinetics of these enzyme-free chemical probes matched that of the gold standard Amplex UltraRed/horseradish peroxidase (HRP) assay, representing a significant advancement in the field and offering a versatile and sensitive tool for studying H<sub>2</sub>O<sub>2</sub>-mediated cell signaling.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5000,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Borinic Acid-Based Fluorogenic Probes as an Alternative to the Amplex Red Assay for Real-Time H<sub>2</sub>O<sub>2</sub> Monitoring in Live Cells.\",\"authors\":\"Mathilde Pucher, Kirrthana Makenthirathasan, Hadrien Jalaber, Thomas LeSaux, Oliver Nüsse, Gilles Doisneau, Yann Bourdreux, Blaise Gatin-Fraudet, Ludovic Jullien, Boris Vauzeilles, Dominique Guianvarc'h, Marie Erard, Dominique Urban\",\"doi\":\"10.1021/acschembio.5c00156\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is a crucial reactive oxygen species (ROS) involved in regulating both physiological and pathological processes. Excessive H<sub>2</sub>O<sub>2</sub> production can lead to oxidative stress, contributing to aging, cancer, and neurodegenerative diseases. In contrast to other ROS exhibiting short lifespans, H<sub>2</sub>O<sub>2</sub> is relatively stable, and its spatial and temporal dynamics are central to understanding its pathophysiological role. Therefore, the development of fluorescent probes that are highly selective, sensitive, and capable of a rapid response is still required. To date, numerous fluorescent probes have been developed. Among them, boronic acid triggers have attracted considerable attention but often suffer from limited reactivity, preventing real-time H<sub>2</sub>O<sub>2</sub> monitoring. To overcome this lack of reactivity, we report the design and synthesis of new borinic acid-based fluorogenic probes for H<sub>2</sub>O<sub>2</sub> detection in cellular environments. These probes are based on a hemicyanine scaffold functionalized with the borinic acid trigger, which demonstrated superior kinetics compared to its boronic counterpart. These probes enable efficient real-time monitoring of H<sub>2</sub>O<sub>2</sub> in cellular models, both extracellularly and intracellularly. The kinetics of these enzyme-free chemical probes matched that of the gold standard Amplex UltraRed/horseradish peroxidase (HRP) assay, representing a significant advancement in the field and offering a versatile and sensitive tool for studying H<sub>2</sub>O<sub>2</sub>-mediated cell signaling.</p>\",\"PeriodicalId\":11,\"journal\":{\"name\":\"ACS Chemical Biology\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2025-06-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Chemical Biology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1021/acschembio.5c00156\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Chemical Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1021/acschembio.5c00156","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Borinic Acid-Based Fluorogenic Probes as an Alternative to the Amplex Red Assay for Real-Time H2O2 Monitoring in Live Cells.
Hydrogen peroxide (H2O2) is a crucial reactive oxygen species (ROS) involved in regulating both physiological and pathological processes. Excessive H2O2 production can lead to oxidative stress, contributing to aging, cancer, and neurodegenerative diseases. In contrast to other ROS exhibiting short lifespans, H2O2 is relatively stable, and its spatial and temporal dynamics are central to understanding its pathophysiological role. Therefore, the development of fluorescent probes that are highly selective, sensitive, and capable of a rapid response is still required. To date, numerous fluorescent probes have been developed. Among them, boronic acid triggers have attracted considerable attention but often suffer from limited reactivity, preventing real-time H2O2 monitoring. To overcome this lack of reactivity, we report the design and synthesis of new borinic acid-based fluorogenic probes for H2O2 detection in cellular environments. These probes are based on a hemicyanine scaffold functionalized with the borinic acid trigger, which demonstrated superior kinetics compared to its boronic counterpart. These probes enable efficient real-time monitoring of H2O2 in cellular models, both extracellularly and intracellularly. The kinetics of these enzyme-free chemical probes matched that of the gold standard Amplex UltraRed/horseradish peroxidase (HRP) assay, representing a significant advancement in the field and offering a versatile and sensitive tool for studying H2O2-mediated cell signaling.
期刊介绍:
ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology.
The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies.
We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.