Deubiquitination Detection of p53 Protein in Living Cells by Fluorescence Cross-Correlation Spectroscopy DOI Creative Commons

Yaoqi Liu,

Chaoqing Dong, Jicun Ren

et al.

ACS Omega, Journal Year: 2023, Volume and Issue: 8(39), P. 36588 - 36596

Published: Sept. 22, 2023

Deubiquitination is a reverse post-translational modification of ubiquitination and plays significant roles in various signal transduction cascades protein stability. The p53 very important tumor-suppressor closely implicates more than 50% human cancers. Although extracellular studies on the deubiquitination were reported, process living cells due to shortage an efficient situ method for single still not clear. In this study, we described studying by combining fluorescence cross-correlation spectroscopy with fluorescent labeling technique. We first constructed stable cell line expressing EGFP-Ub-p53-mCherry as substrate deubiquitination. Then, established monitoring cells. Based amplitudes correlation curves from cells, obtained percentage evaluating level Furthermore, studied effects ubiquitin structures found that C-terminal Gly75-Gly76 motif key location cannot occur when lacks motif. Our results documented developed strategy study proteins

Language: Английский

Deubiquitination Detection of p53 Protein in Living Cells by Fluorescence Cross-Correlation Spectroscopy DOI Creative Commons

Yaoqi Liu,

Chaoqing Dong, Jicun Ren

et al.

ACS Omega, Journal Year: 2023, Volume and Issue: 8(39), P. 36588 - 36596

Published: Sept. 22, 2023

Deubiquitination is a reverse post-translational modification of ubiquitination and plays significant roles in various signal transduction cascades protein stability. The p53 very important tumor-suppressor closely implicates more than 50% human cancers. Although extracellular studies on the deubiquitination were reported, process living cells due to shortage an efficient situ method for single still not clear. In this study, we described studying by combining fluorescence cross-correlation spectroscopy with fluorescent labeling technique. We first constructed stable cell line expressing EGFP-Ub-p53-mCherry as substrate deubiquitination. Then, established monitoring cells. Based amplitudes correlation curves from cells, obtained percentage evaluating level Furthermore, studied effects ubiquitin structures found that C-terminal Gly75-Gly76 motif key location cannot occur when lacks motif. Our results documented developed strategy study proteins

Language: Английский

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