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Although CDK6 makes an attractive cancer biomarker and target, there are no means of assessing its activity in a complex environment. In this study, we describe the design, engineering and characterisation of a fluorescent peptide biosensor derived from 6\u2010phosphofructokinase that reports on CDK6 kinase activity through sensitive changes in fluorescence intensity. This biosensor can report on CDK6 activity in a dose\u2010dependent fashion, thereby enabling quantification of differences in kinase activity in complex and physiologically relevant environments. Further implementation of this biosensor in different lung and melanoma cell lines, as well as in mesothelioma cell lines derived from patients together with a CDK4 biosensor highlighted differences in kinase activity between CDK6 and CDK4 kinase. This work demonstrates the utility of these selective tools for monitoring two closely related kinases comparatively and simultaneously in the same samples, thereby offering attractive perspectives for diagnostic and therapeutic purposes.<\/jats:p>","DOI":"10.1002\/cbic.202000677","type":"journal-article","created":{"date-parts":[[2020,10,28]],"date-time":"2020-10-28T06:47:33Z","timestamp":1603867653000},"page":"1065-1071","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Fluorescent Peptide Biosensor for Probing CDK6 Kinase Activity in Lung Cancer Cell Extracts"],"prefix":"10.1002","volume":"22","author":[{"given":"Jessica","family":"Soamalala","sequence":"first","affiliation":[{"name":"Institut des Biomol\u00e9cules Max Mousseron-IBMM-UMR 5247 Universit\u00e9 de Montpellier, Facult\u00e9 de Pharmacie  15, Av. Charles Flahault 34093 Montpellier France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebastien","family":"Diot","sequence":"additional","affiliation":[{"name":"Institut des Biomol\u00e9cules Max Mousseron-IBMM-UMR 5247 Universit\u00e9 de Montpellier, Facult\u00e9 de Pharmacie  15, Av. Charles Flahault 34093 Montpellier France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgan","family":"Pellerano","sequence":"additional","affiliation":[{"name":"Institut des Biomol\u00e9cules Max Mousseron-IBMM-UMR 5247 Universit\u00e9 de Montpellier, Facult\u00e9 de Pharmacie  15, Av. 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