An in vitro Förster resonance energy transfer-based high-throughput screening assay identifies inhibitors of SUMOylation E2 Ubc9
SUMOylation is one of the posttranslational modifications that mediate cellular activities such as transcription, DNA repair, and signal transduction and is involved in the cell cycle. However, only a limited number of small molecule inhibitors have been identified to study its role in cellular processes. Here, we report a Förster resonance energy transfer (FRET) high-throughput screening assay based on the interaction between E2 Ubc9 and E3 PIAS1. Of the 3200 compounds screened, 34 (1.1%) showed higher than 50% inhibition and 4 displayed dose-response inhibitory effects. By combining this method with a label-free surface plasmon resonance (SPR) assay, false positives were excluded leading to discovering WNN0605-F008 and WNN1062-D002 that bound to Ubc9 with KD values of 1.93 ± 0.62 and 5.24 ± 3.73 μM, respectively. We examined the effect of the two compounds on SUMO2-mediated SUMOylation of RanGAP1, only WNN0605-F008 significantly inhibited RanGAP1 SUMOylation, whereas WNN1062-D002 did not show any inhibition. These compounds, with novel chemical scaffolds, may serve as the initial material for developing new SUMOylation inhibitors.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:41 |
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| Enthalten in: |
Acta pharmacologica Sinica - 41(2020), 11 vom: 27. Nov., Seite 1497-1506 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wang, Yu-Zhe [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 07.07.2021 Date Revised 05.10.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1038/s41401-020-0405-7 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM309254957 |
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| 100 | 1 | |a Wang, Yu-Zhe |e verfasserin |4 aut | |
| 245 | 1 | 3 | |a An in vitro Förster resonance energy transfer-based high-throughput screening assay identifies inhibitors of SUMOylation E2 Ubc9 |
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| 500 | |a Citation Status MEDLINE | ||
| 520 | |a SUMOylation is one of the posttranslational modifications that mediate cellular activities such as transcription, DNA repair, and signal transduction and is involved in the cell cycle. However, only a limited number of small molecule inhibitors have been identified to study its role in cellular processes. Here, we report a Förster resonance energy transfer (FRET) high-throughput screening assay based on the interaction between E2 Ubc9 and E3 PIAS1. Of the 3200 compounds screened, 34 (1.1%) showed higher than 50% inhibition and 4 displayed dose-response inhibitory effects. By combining this method with a label-free surface plasmon resonance (SPR) assay, false positives were excluded leading to discovering WNN0605-F008 and WNN1062-D002 that bound to Ubc9 with KD values of 1.93 ± 0.62 and 5.24 ± 3.73 μM, respectively. We examined the effect of the two compounds on SUMO2-mediated SUMOylation of RanGAP1, only WNN0605-F008 significantly inhibited RanGAP1 SUMOylation, whereas WNN1062-D002 did not show any inhibition. These compounds, with novel chemical scaffolds, may serve as the initial material for developing new SUMOylation inhibitors | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Förster resonance energy transfer | |
| 650 | 4 | |a PIAS1 | |
| 650 | 4 | |a SUMOylation inhibitor | |
| 650 | 4 | |a Ubc9 | |
| 650 | 4 | |a WNN0605-F008 | |
| 650 | 4 | |a high-throughput screening | |
| 650 | 4 | |a surface plasmon resonance | |
| 650 | 7 | |a Enzyme Inhibitors |2 NLM | |
| 650 | 7 | |a GTPase-Activating Proteins |2 NLM | |
| 650 | 7 | |a PIAS1 protein, human |2 NLM | |
| 650 | 7 | |a Protein Inhibitors of Activated STAT |2 NLM | |
| 650 | 7 | |a RANGAP1 protein, human |2 NLM | |
| 650 | 7 | |a Small Molecule Libraries |2 NLM | |
| 650 | 7 | |a Small Ubiquitin-Related Modifier Proteins |2 NLM | |
| 650 | 7 | |a Ubiquitin-Conjugating Enzymes |2 NLM | |
| 650 | 7 | |a EC 2.3.2.23 |2 NLM | |
| 650 | 7 | |a ubiquitin-conjugating enzyme UBC9 |2 NLM | |
| 650 | 7 | |a EC 6.3.2.- |2 NLM | |
| 700 | 1 | |a Liu, Xiao |e verfasserin |4 aut | |
| 700 | 1 | |a Way, George |e verfasserin |4 aut | |
| 700 | 1 | |a Madarha, Vipul |e verfasserin |4 aut | |
| 700 | 1 | |a Zhou, Qing-Tong |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, De-Hua |e verfasserin |4 aut | |
| 700 | 1 | |a Liao, Jia-Yu |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Ming-Wei |e verfasserin |4 aut | |
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