Comparative Analysis of a FRET-based PLK1 Kinase Assay to Identify PLK1 inhibitors for Chemotherapy
- Authors
- Shin, Sol-Bi; Woo, Sang-Uk; Lee, Young-Joo; Yim, Hyungshin
- Issue Date
- Mar-2017
- Publisher
- INT INST ANTICANCER RESEARCH
- Keywords
- PLK1; FRET; radioisotope; immunoblot; BI 2536; kinase assay
- Citation
- ANTICANCER RESEARCH, v.37, no.3, pp.1177 - 1183
- Indexed
- SCIE
SCOPUS
- Journal Title
- ANTICANCER RESEARCH
- Volume
- 37
- Number
- 3
- Start Page
- 1177
- End Page
- 1183
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/10124
- DOI
- 10.21873/anticanres.11431
- ISSN
- 0250-7005
- Abstract
- Advanced techniques for detecting kinase inhibitors are in demand due to limitations of traditional approaches. Here, we used a fluorescence resonance energy transfer (FRET)-based kinase assay, a sensitive fluorescence turn-on biosensing platform, to identify a Polo-like kinase 1 (PLK1) inhibitor. The assay was developed with the Z '- Lyte(TM) FRET-peptide and PLK1 kinase purified from a baculovirus expression system. Using PLK1 inhibitors, sensitivity and efficiency of this FRET-based PLK1 kinase assay were compared to those of radioisotope-based and immunoblot-based assays. Although the inhibitory activity of BI 2536 against PLK1 kinase in each assay was almost the same, the FRET-based PLK1 kinase assay was much easier, faster, safer, and more convenient than a radioisotope-based assay or an immunoblot-based traditional kinase assay. From our findings, we suggest that a FRET-based PLK1 kinase assay is an advanced tool which overcomes the limitations of previous traditional kinase assays to detect kinase inhibitors for the development of anticancer drugs.
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