Development of a kind of RG108-Fluorescein conjugates for detection of DNA methyltransferase 1 (DNMT1) in living cells
| Autor: | Chaochao Gong, Qian Zhang, Minli Ruan, Ling Xu, Zhonglian Cao, Cheng Qunxian |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
DNA (Cytosine-5-)-Methyltransferase 1
Biophysics Uterine Cervical Neoplasms Phthalimides environment and public health 01 natural sciences Biochemistry DNA methyltransferase HeLa 03 medical and health sciences chemistry.chemical_compound medicine Humans Surface plasmon resonance Binding site Fluorescein Molecular Biology 030304 developmental biology Fluorescent Dyes 0303 health sciences biology urogenital system Chemistry 010401 analytical chemistry Tryptophan Cell Biology DNA Methylation biology.organism_classification 0104 chemical sciences Molecular Docking Simulation medicine.anatomical_structure embryonic structures DNA methylation DNMT1 Female Nucleus Protein Processing Post-Translational HeLa Cells Protein Binding |
| Zdroj: | Analytical biochemistry. 607 |
| ISSN: | 1096-0309 |
| Popis: | DNA methyltransferase 1 (DNMT1) is one of the most essential proteins in propagating DNA methylation patterns during replication. Developing methods to assess the expression level of DNMT1 will enable study of gene methylation abnormalities. Thus, a series of fluorescein-conjugated RG108 derivatives were designed and synthesized in the current study. The affinity of the derivatives with DNMT1 was evaluated using surface plasmon resonance. Permeability of the derivatives through the cytomembrane and nuclear envelope was evaluated via confocal imaging. Probe 8a was found to compete with RG108 binding to DNMT1 in the nucleus of HeLa cells, suggesting that probe 8a and RG108 share the same binding site. A HeLa cell model with 4.05-fold overexpression of DNMT1 was constructed and used to evaluate probe 8a. Probe 8a was found to be significantly increased in the nucleus of DNMT1 overexpressing cells. These results indicate that fluorescent probes derived from RG108 have the potential to be used for evaluating the expression level of DNMT1 in living cells. |
| Databáze: | OpenAIRE |
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