Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells
PLK1 is a protein kinase that regulates mitosis and is both an important oncology drug target and a potential antitarget of drugs for the DNA damage response pathway or anti-infective host kinases. To expand the range of live cell NanoBRET target engagement assays to include PLK1, we developed an en...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/7/2950 |
| _version_ | 1797607447563599872 |
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| author | Xuan Yang Jeffery L. Smith Michael T. Beck Jennifer M. Wilkinson Ani Michaud James D. Vasta Matthew B. Robers Timothy M. Willson |
| author_facet | Xuan Yang Jeffery L. Smith Michael T. Beck Jennifer M. Wilkinson Ani Michaud James D. Vasta Matthew B. Robers Timothy M. Willson |
| author_sort | Xuan Yang |
| collection | DOAJ |
| description | PLK1 is a protein kinase that regulates mitosis and is both an important oncology drug target and a potential antitarget of drugs for the DNA damage response pathway or anti-infective host kinases. To expand the range of live cell NanoBRET target engagement assays to include PLK1, we developed an energy transfer probe based on the anilino-tetrahydropteridine chemotype found in several selective PLK inhibitors. Probe <b>11</b> was used to configure NanoBRET target engagement assays for PLK1, PLK2, and PLK3 and measure the potency of several known PLK inhibitors. In-cell target engagement for PLK1 was in good agreement with the reported cellular potency for the inhibition of cell proliferation. Probe <b>11</b> enabled the investigation of the promiscuity of adavosertib, which had been described as a dual PLK1/WEE1 inhibitor in biochemical assays. Live cell target engagement analysis of adavosertib via NanoBRET demonstrated PLK activity at micromolar concentrations but only selective engagement of WEE1 at clinically relevant doses. |
| first_indexed | 2024-03-11T05:30:09Z |
| format | Article |
| id | doaj.art-d38f4c63c4d142138e67e669810bc196 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-11T05:30:09Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-d38f4c63c4d142138e67e669810bc1962023-11-17T17:11:20ZengMDPI AGMolecules1420-30492023-03-01287295010.3390/molecules28072950Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live CellsXuan Yang0Jeffery L. Smith1Michael T. Beck2Jennifer M. Wilkinson3Ani Michaud4James D. Vasta5Matthew B. Robers6Timothy M. Willson7Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAStructural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAPromega Corporation, 2800 Woods Hollow Road, Madison, WI 53719, USAPromega Corporation, 2800 Woods Hollow Road, Madison, WI 53719, USAPromega Corporation, 2800 Woods Hollow Road, Madison, WI 53719, USAPromega Corporation, 2800 Woods Hollow Road, Madison, WI 53719, USAPromega Corporation, 2800 Woods Hollow Road, Madison, WI 53719, USAStructural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAPLK1 is a protein kinase that regulates mitosis and is both an important oncology drug target and a potential antitarget of drugs for the DNA damage response pathway or anti-infective host kinases. To expand the range of live cell NanoBRET target engagement assays to include PLK1, we developed an energy transfer probe based on the anilino-tetrahydropteridine chemotype found in several selective PLK inhibitors. Probe <b>11</b> was used to configure NanoBRET target engagement assays for PLK1, PLK2, and PLK3 and measure the potency of several known PLK inhibitors. In-cell target engagement for PLK1 was in good agreement with the reported cellular potency for the inhibition of cell proliferation. Probe <b>11</b> enabled the investigation of the promiscuity of adavosertib, which had been described as a dual PLK1/WEE1 inhibitor in biochemical assays. Live cell target engagement analysis of adavosertib via NanoBRET demonstrated PLK activity at micromolar concentrations but only selective engagement of WEE1 at clinically relevant doses.https://www.mdpi.com/1420-3049/28/7/2950kinaseassaydrugin celltarget engagement |
| spellingShingle | Xuan Yang Jeffery L. Smith Michael T. Beck Jennifer M. Wilkinson Ani Michaud James D. Vasta Matthew B. Robers Timothy M. Willson Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells Molecules kinase assay drug in cell target engagement |
| title | Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells |
| title_full | Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells |
| title_fullStr | Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells |
| title_full_unstemmed | Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells |
| title_short | Development of Cell Permeable NanoBRET Probes for the Measurement of PLK1 Target Engagement in Live Cells |
| title_sort | development of cell permeable nanobret probes for the measurement of plk1 target engagement in live cells |
| topic | kinase assay drug in cell target engagement |
| url | https://www.mdpi.com/1420-3049/28/7/2950 |
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