Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine
Over the last two decades, the development of fluorescent probes has transformed the way of measuring physiological parameters in intact cells, including in the field of redox biology. We developed a genetically encoded biosensor called CyReB to monitor intracellular cysteine in real time. This bios...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | Redox Biology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231725002988 |
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| author | Damien Caubrière Arthur de Butler Anna Moseler Pauline Leverrier Jean-François Collet Andreas J. Meyer Nicolas Rouhier Jérémy Couturier |
| author_facet | Damien Caubrière Arthur de Butler Anna Moseler Pauline Leverrier Jean-François Collet Andreas J. Meyer Nicolas Rouhier Jérémy Couturier |
| author_sort | Damien Caubrière |
| collection | DOAJ |
| description | Over the last two decades, the development of fluorescent probes has transformed the way of measuring physiological parameters in intact cells, including in the field of redox biology. We developed a genetically encoded biosensor called CyReB to monitor intracellular cysteine in real time. This biosensor exploits the ability of a particular bacterial cysteine desulfurase to promote the oxidation of reduction-oxidation-sensitive green fluorescent protein 2 in the presence of cysteine. The specificity, sensitivity, and the oxidation-reduction dynamics of CyReB were first investigated in vitro before its in vivo functionality was confirmed by expressing CyReB in Escherichia coli and Saccharomyces cerevisiae cells. Expressing CyReB or an inactive version in wild-type and various mutant strains of Escherichia coli showed that this sensor could be used to monitor intracellular cysteine dynamics, particularly in the context of the cysteine-cystine shuttle system. This work demonstrates how using this cysteine biosensor should provide new insights into the metabolism of cysteine and cysteine-related pathways in various model organisms. |
| format | Article |
| id | doaj-art-390af1349b684ce2b2cc966fbf7d24cf |
| institution | Kabale University |
| issn | 2213-2317 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj-art-390af1349b684ce2b2cc966fbf7d24cf2025-08-24T05:12:39ZengElsevierRedox Biology2213-23172025-09-018510378510.1016/j.redox.2025.103785Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteineDamien Caubrière0Arthur de Butler1Anna Moseler2Pauline Leverrier3Jean-François Collet4Andreas J. Meyer5Nicolas Rouhier6Jérémy Couturier7Université de Lorraine, INRAE, IAM, F-54000, Nancy, FranceUniversité de Lorraine, INRAE, IAM, F-54000, Nancy, FranceInstitute of Crop Science and Resource Conservation (INRES) - Chemical Signalling, University of Bonn, 53117, Bonn, GermanyWELBIO Department, WEL Research Institute, Avenue Pasteur 6, 1300, Wavre, Belgium; de Duve Institute, Université Catholique de Louvain, Avenue Hippocrate 75, 1200, Brussels, BelgiumWELBIO Department, WEL Research Institute, Avenue Pasteur 6, 1300, Wavre, Belgium; de Duve Institute, Université Catholique de Louvain, Avenue Hippocrate 75, 1200, Brussels, BelgiumInstitute of Crop Science and Resource Conservation (INRES) - Chemical Signalling, University of Bonn, 53117, Bonn, GermanyUniversité de Lorraine, INRAE, IAM, F-54000, Nancy, France; Institut Universitaire de France, F-75000, Paris, FranceUniversité de Lorraine, INRAE, IAM, F-54000, Nancy, France; Institut Universitaire de France, F-75000, Paris, France; Corresponding author. Université de Lorraine, Interactions Arbres/Microorganismes, UMR1136, Vandœuvre-lès-Nancy, F-54500, France.Over the last two decades, the development of fluorescent probes has transformed the way of measuring physiological parameters in intact cells, including in the field of redox biology. We developed a genetically encoded biosensor called CyReB to monitor intracellular cysteine in real time. This biosensor exploits the ability of a particular bacterial cysteine desulfurase to promote the oxidation of reduction-oxidation-sensitive green fluorescent protein 2 in the presence of cysteine. The specificity, sensitivity, and the oxidation-reduction dynamics of CyReB were first investigated in vitro before its in vivo functionality was confirmed by expressing CyReB in Escherichia coli and Saccharomyces cerevisiae cells. Expressing CyReB or an inactive version in wild-type and various mutant strains of Escherichia coli showed that this sensor could be used to monitor intracellular cysteine dynamics, particularly in the context of the cysteine-cystine shuttle system. This work demonstrates how using this cysteine biosensor should provide new insights into the metabolism of cysteine and cysteine-related pathways in various model organisms.http://www.sciencedirect.com/science/article/pii/S2213231725002988Cysteine desulfuraseCysteineCystineFluorescent biosensorroGFP2 |
| spellingShingle | Damien Caubrière Arthur de Butler Anna Moseler Pauline Leverrier Jean-François Collet Andreas J. Meyer Nicolas Rouhier Jérémy Couturier Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine Redox Biology Cysteine desulfurase Cysteine Cystine Fluorescent biosensor roGFP2 |
| title | Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine |
| title_full | Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine |
| title_fullStr | Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine |
| title_full_unstemmed | Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine |
| title_short | Fusion of a bacterial cysteine desulfurase to redox-sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine |
| title_sort | fusion of a bacterial cysteine desulfurase to redox sensitive green fluorescent protein produces a highly sensitive cysteine biosensor for monitoring changes in intracellular cysteine |
| topic | Cysteine desulfurase Cysteine Cystine Fluorescent biosensor roGFP2 |
| url | http://www.sciencedirect.com/science/article/pii/S2213231725002988 |
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