A Fluorophore‐labeled Peptide of Human Ribosomal Protein <scp>S3</scp> for the Detection of the 8‐Oxoguanine within the Cells

Han, Se Hee; Hahm, Soo-Hyun; Tran, An Hue Vy; Park, Jin Woo; Chung, Ji Hyung; Park, Geon Tae; Han, Ye

doi:10.1002/bkcs.10472

Cited by 4 publications

(8 citation statements)

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“…In a previous study, we confirmed that an S3 probe containing a K132 (K134 of Drosophila melanogaster RpS3) amino acid residue, which plays a crucial role in the binding of hrpS3 and 8-oxoG, specifically binds to substrates containing 8-oxoG (38). In addition to 8-oxoG, hRpS3 is also known to have binding affinity to AP sites, and K132 amino acid residues are known to play an important role in specific binding to AP sites (43,46).…”

Section: Binding Of the Tat-s3 Peptide Probe To Substrate Containing supporting

confidence: 67%

“…We previously developed an imaging probe to detect 8-oxoG using a specific peptide of hRpS3 (38). A TAT (47-57, YGRKKRRQRRR) peptide that can penetrate the cell was attached at the C-terminal of the S3 probe and a two amino acid GG linker was added between the S3 probe and TAT in designing the new TAT-S3 probe.…”

Section: Synthesis Of the Tat-s3 Peptide Probementioning

confidence: 99%

“…In previous studies, we generated an hRpS3-peptide probe that could be used to detect 8-oxoG via a fluorescence assay (38). We generated a new probe for 8-oxoG and AP sites consisting of TAT peptide and hRpS3.…”

mentioning

confidence: 99%

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Detection of 8-oxoguanine and Apurinic/Apyrimidinic Sites Using a Fluorophore-Labeled Probe With Cell-penetrating Ability

Kang

Shin

Kim

et al. 2019

Preprint

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Background Detection and quantification of 8-oxo-7,8-dihydroguanine (8-oxoG) within cells are important for studying the molecular mechanisms of cancer. Human ribosomal protein S3 (hRpS3), which is involved in DNA repair, has a high binding affinity to 8-oxoG. We developed an imaging probe to detect 8-oxoG using a specific peptide from hRpS3. Transactivator (TAT) proteins are commonly known to have cell-penetrating properties. Therefore, we developed the TAT-S3 probe via attaching a TAT peptide to our imaging probe. Result A DNA binding assay was conducted to confirm that our probe bound to 8-oxoG and AP sites. We confirmed that the TAT-S3 probe is present in the mitochondria, without permeabilization, and the TAT-S3 probe also showed fluorescence in H2O2-treated HeLa cells and zebrafish embryos. Treatment with MitoQ, a mitochondria-targeted anti-oxidant, reduced TAT-S3 probe fluorescence. Additionally, treatment with O8, an inhibitor of OGG1, increased probe fluorescence. A competition assay was conducted to utilize an aldehyde reaction probe (ARP) and methoxyamine (MX) to confirm the binding of the TAT-S3 to the AP sites. The TAT-S3 probe showed binding to AP sites competitive with ARP and MX. Conclusion These results revealed that the TAT-S3 probe successfully detected the presence of 8-oxoG and AP sites in damaged cells. The TAT-S3 probe may have applications for the detection of diseases caused by reactive oxygen species (ROS).

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Section: Binding Of the Tat-s3 Peptide Probe To Substrate Containing supporting

confidence: 67%

Section: Synthesis Of the Tat-s3 Peptide Probementioning

confidence: 99%

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Detection of 8-oxoguanine and Apurinic/Apyrimidinic Sites Using a Fluorophore-Labeled Probe With Cell-penetrating Ability

Kang

Shin

Kim

et al. 2019

Preprint

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“…1 However, the genomic DNAs are frequently damaged by various exogenous and endogenous factors, creating $10 5 lesions (e.g., modied bases, abasic sites, DNA adducts, DNA strand breaks, intra-and inter-strand crosslinks) in a single cell per day. 2,3 Persistent DNA damage may severely induce base substitutions, insertions, deletions and chromosomal rearrangements, causing genomic instability, premature ageing, developmental disorders and carcinogenesis progression. 4,5 Cells have evolved multiple protection mechanisms to specially repair a wide range of DNA lesions.…”

Section: Introductionmentioning

confidence: 99%

Construction of a self-directed replication system for label-free and real-time sensing of repair glycosylases with zero background

2020

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“…In previous studies, we generated an hRpS3-peptide probe that could be used to detect 8-oxoG via a fluorescence assay [37]. We generated a new probe for 8-oxoG and AP sites consisting of TAT peptide and hRpS3, termed TAT-S3.…”

Section: Introductionmentioning

confidence: 99%

Detection of 8-oxoguanine and apurinic/apyrimidinic sites using a fluorophore-labeled probe with cell-penetrating ability

Kang

Shin

Kim

et al. 2019

BMC Mol and Cell Biol

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BackgroundReactive oxygen species (ROS) produce different lesions in DNA by ROS-induced DNA damage. Detection and quantification of 8-oxo-7,8-dihydroguanine (8-oxoG) within cells are important for study. Human ribosomal protein S3 (hRpS3) has a high binding affinity to 8-oxoG. In this study, we developed an imaging probe to detect 8-oxoG using a specific peptide from hRpS3. Transactivator (TAT) proteins are known to have cell-penetrating properties. Therefore, we developed a TAT-S3 probe by attaching a TAT peptide to our imaging probe.ResultsA DNA binding assay was conducted to confirm that our probe bound to 8-oxoG and apurinic/apyrimidinic (AP) sites. We confirmed that the TAT-S3 probe localized in the mitochondria, without permeabilization, and fluoresced in H2O2-treated HeLa cells and zebrafish embryos. Treatment with Mitoquinone (MitoQ), a mitochondria-targeted antioxidant, reduced TAT-S3 probe fluorescence. Additionally, treatment with O8, an inhibitor of OGG1, increased probe fluorescence. A competition assay was conducted with an aldehyde reaction probe (ARP) and methoxyamine (MX) to confirm binding of TAT-S3 to the AP sites. The TAT-S3 probe showed competitive binding to AP sites with ARP and MX.ConclusionsThese results revealed that the TAT-S3 probe successfully detected the presence of 8-oxoG and AP sites in damaged cells. The TAT-S3 probe may have applications for the detection of diseases caused by reactive oxygen species.

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A Fluorophore‐labeled Peptide of Human Ribosomal Protein S3 for the Detection of the 8‐Oxoguanine within the Cells

Cited by 4 publications

References 41 publications

Detection of 8-oxoguanine and Apurinic/Apyrimidinic Sites Using a Fluorophore-Labeled Probe With Cell-penetrating Ability

Detection of 8-oxoguanine and Apurinic/Apyrimidinic Sites Using a Fluorophore-Labeled Probe With Cell-penetrating Ability

Construction of a self-directed replication system for label-free and real-time sensing of repair glycosylases with zero background

Detection of 8-oxoguanine and apurinic/apyrimidinic sites using a fluorophore-labeled probe with cell-penetrating ability

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