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Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species

The intracellular concentrations of oxygen and reactive oxygen species (ROS) in living cells represent critical information for investigating physiological and pathological conditions. Real-time measurement often relies on genetically encoded proteins that are responsive to fluctuations in either ox...

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Autores principales: Marchetti, Marialaura, Ronda, Luca, Cozzi, Monica, Bettati, Stefano, Bruno, Stefano
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10611200/
https://www.ncbi.nlm.nih.gov/pubmed/37896609
http://dx.doi.org/10.3390/s23208517
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author Marchetti, Marialaura
Ronda, Luca
Cozzi, Monica
Bettati, Stefano
Bruno, Stefano
author_facet Marchetti, Marialaura
Ronda, Luca
Cozzi, Monica
Bettati, Stefano
Bruno, Stefano
author_sort Marchetti, Marialaura
collection PubMed
description The intracellular concentrations of oxygen and reactive oxygen species (ROS) in living cells represent critical information for investigating physiological and pathological conditions. Real-time measurement often relies on genetically encoded proteins that are responsive to fluctuations in either oxygen or ROS concentrations. The direct binding or chemical reactions that occur in their presence either directly alter the fluorescence properties of the binding protein or alter the fluorescence properties of fusion partners, mostly consisting of variants of the green fluorescent protein. Oxygen sensing takes advantage of several mechanisms, including (i) the oxygen-dependent hydroxylation of a domain of the hypoxia-inducible factor-1, which, in turn, promotes its cellular degradation along with fluorescent fusion partners; (ii) the naturally oxygen-dependent maturation of the fluorophore of green fluorescent protein variants; and (iii) direct oxygen binding by proteins, including heme proteins, expressed in fusion with fluorescent partners, resulting in changes in fluorescence due to conformational alterations or fluorescence resonance energy transfer. ROS encompass a group of highly reactive chemicals that can interconvert through various chemical reactions within biological systems, posing challenges for their selective detection through genetically encoded sensors. However, their general reactivity, and particularly that of the relatively stable oxygen peroxide, can be exploited for ROS sensing through different mechanisms, including (i) the ROS-induced formation of disulfide bonds in engineered fluorescent proteins or fusion partners of fluorescent proteins, ultimately leading to fluorescence changes; and (ii) conformational changes of naturally occurring ROS-sensing domains, affecting the fluorescence properties of fusion partners. In this review, we will offer an overview of these genetically encoded biosensors.
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spelling pubmed-106112002023-10-28 Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species Marchetti, Marialaura Ronda, Luca Cozzi, Monica Bettati, Stefano Bruno, Stefano Sensors (Basel) Review The intracellular concentrations of oxygen and reactive oxygen species (ROS) in living cells represent critical information for investigating physiological and pathological conditions. Real-time measurement often relies on genetically encoded proteins that are responsive to fluctuations in either oxygen or ROS concentrations. The direct binding or chemical reactions that occur in their presence either directly alter the fluorescence properties of the binding protein or alter the fluorescence properties of fusion partners, mostly consisting of variants of the green fluorescent protein. Oxygen sensing takes advantage of several mechanisms, including (i) the oxygen-dependent hydroxylation of a domain of the hypoxia-inducible factor-1, which, in turn, promotes its cellular degradation along with fluorescent fusion partners; (ii) the naturally oxygen-dependent maturation of the fluorophore of green fluorescent protein variants; and (iii) direct oxygen binding by proteins, including heme proteins, expressed in fusion with fluorescent partners, resulting in changes in fluorescence due to conformational alterations or fluorescence resonance energy transfer. ROS encompass a group of highly reactive chemicals that can interconvert through various chemical reactions within biological systems, posing challenges for their selective detection through genetically encoded sensors. However, their general reactivity, and particularly that of the relatively stable oxygen peroxide, can be exploited for ROS sensing through different mechanisms, including (i) the ROS-induced formation of disulfide bonds in engineered fluorescent proteins or fusion partners of fluorescent proteins, ultimately leading to fluorescence changes; and (ii) conformational changes of naturally occurring ROS-sensing domains, affecting the fluorescence properties of fusion partners. In this review, we will offer an overview of these genetically encoded biosensors. MDPI 2023-10-17 /pmc/articles/PMC10611200/ /pubmed/37896609 http://dx.doi.org/10.3390/s23208517 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Review
Marchetti, Marialaura
Ronda, Luca
Cozzi, Monica
Bettati, Stefano
Bruno, Stefano
Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species
title Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species
title_full Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species
title_fullStr Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species
title_full_unstemmed Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species
title_short Genetically Encoded Biosensors for the Fluorescence Detection of O(2) and Reactive O(2) Species
title_sort genetically encoded biosensors for the fluorescence detection of o(2) and reactive o(2) species
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10611200/
https://www.ncbi.nlm.nih.gov/pubmed/37896609
http://dx.doi.org/10.3390/s23208517
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