A guide to genetically encoded tools for the study of H<sub>2</sub>O<sub>2</sub>

Smolyarova, Daria D.; Podgorny, Oleg V.; Bilan, Dmitry S.; Belousov, Vsevolod V.

doi:10.1111/febs.16088

Cited by 35 publications

(19 citation statements)

References 125 publications

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“…This disagreement suggests that the role of the active sites in the form of V ). This is consistent with the observation that H 2 O 2 is the most longlived ROS 101 and so induces considerable cytotoxicity on cancer cells. 102,103 However, the effect of the medium is clear in that the cell viability at 48 h decreased to 35 ± 2%.…”

Section: Cytotoxicity Of Nanoceria Against Tumor Cells and Normal Cellssupporting

confidence: 92%

ROS‐mediated anticancer effects of EGFR‐targeted nanoceria

Johnson,

Koshy,

Kopecky

et al. 2023

J Biomedical Materials Res

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The therapeutic effectiveness of anticancer drugs, including nanomedicines, can be enhanced with active receptor‐targeting strategies. Epidermal growth factor receptor (EGFR) is an important cancer biomarker, constitutively expressed in sarcoma patients of different histological types. The present work reports materials and in vitro biomedical analyses of silanized (passive delivery) and/or EGF‐functionalized (active delivery) ceria nanorods exhibiting highly defective catalytically active surfaces. The EGFR‐targeting efficiency of nanoceria was confirmed by receptor‐binding studies. Increased cytotoxicity and reactive oxygen species (ROS) production were observed for EGF‐functionalized nanoceria owing to enhanced cellular uptake by HT‐1080 fibrosarcoma cells. The uptake was confirmed by TEM and confocal microscopy. Silanized nanoceria demonstrated negligible/minimal cytotoxicity toward healthy MRC‐5 cells at 24 and 48 h, whereas this was significant at 72 h owing to a nanoceria accumulation effect. In contrast, considerable cytotoxicity toward the cancer cells was exhibited at all three times points. The ROS generation and associated cytotoxicity were moderated by the equilibrium between catalysis by ceria, generation of cell debris, and blockage of active sites. EGFR‐targeting is shown to enhance the uptake levels of nanoceria by cancer cells, subsequently enhancing the overall anticancer activity and therapeutic performance of ceria.

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Section: Cytotoxicity Of Nanoceria Against Tumor Cells and Normal Cellssupporting

confidence: 92%

ROS‐mediated anticancer effects of EGFR‐targeted nanoceria

Johnson,

Koshy,

Kopecky

et al. 2023

J Biomedical Materials Res

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“…The CCK8 results demonstrated that tetrapeptide exhibited good biocompatibility even at the concentration 6 mg/mL (Figure c). One of the major advantages of mimetic tetrapeptide is the ease of use without a priori genetic manipulations . To demonstrate the suitability of the CFFH tetrapeptide for detecting hydrogen peroxide in acidic organelles, we pretreated the HeLa cells with tetrapeptide for 24 h. The confocal laser scanning microscopy (CLSM) revealed that weak blue fluorescence and a visible green spot were observed in the cells, which suggested that the roGFP2 mimetic tetrapeptide could monitor hydrogen peroxide in an acidic microenvironment (Figure d).…”

Section: Resultsmentioning

confidence: 99%

Rethreading Design of Ratiometric roGFP2 Mimetic Peptide for Hydrogen Peroxide Sensing

Kong

et al. 2023

Anal. Chem.

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Reconstruction of the miniaturized peptide to mimic the tailored functions of protein has been attractive but challenging. Herein, initialized from the crystal structure of redox-sensitive green fluorescent protein-2 (roGFP2), we propose a practical approach to construct the roGFP2 mimetic peptide by rethreading the aromatic residues adjacent to the chromophore fragment. By fine-tuning the residues of peptides, a mini tetrapeptide (Cys-Phe-Phe-His) was designed, which can act as a hydrogen peroxide sensor using its ratiometric fluorescence. The roGFP2 mimetic tetrapeptide is biocompatible and photostable and has competitive fluorescent properties with roGFP2 by the virtue of its assembly induced emissions. We expand the ratiometric tetrapeptide for sensing hydrogen peroxide in acidic chambers. The results provide a promising approach for the artificial design of miniaturized peptides with the desired function.

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“…Later, several H 2 O 2 sensors were developed by fusing roGFP or rxYFP to thiol-based peroxidases, such as Orp1, 389 Tsa2, 390 Prx1 391 and Tpx1. 392,393 The peroxidases oxidise the cysteine pair on the roGFP later react with H 2 O 2 .…”

Section: Genetically Encoded Fluorescent Probesmentioning

confidence: 99%