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Multiplexable fluorescence lifetime imaging (FLIM) probes for Abl and Src-family kinases.

ABSTRACT: Many commonly employed strategies to map kinase activities in live cells require expression of genetically encoded proteins (e.g. FRET sensors). In this work, we describe the development and preliminary application of a set of cell-penetrating, fluorophore labelled peptide substrates for fluorescence lifetime imaging (FLIM) of Abl and Src-family kinase activities. These probes do not rely on FRET pairs or genetically-encoded protein expression. We further demonstrate probe multiplexing and pixel-by-pixel quantification to estimate the relative proportion of modified probe, suggesting that this strategy will be useful for detailed mapping of single cell and subcellular dynamics of multiple kinases concurrently in live cells.

SUBMITTER: Jena S

PROVIDER: S-EPMC7679178 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Multiplexable fluorescence lifetime imaging (FLIM) probes for Abl and Src-family kinases.

Jena Sampreeti S Damayanti Nur P NP Tan Jackie J Pratt Erica D ED Irudayaraj Joseph M K JMK Parker Laurie L LL

Chemical communications (Cambridge, England) 20201009 87

Many commonly employed strategies to map kinase activities in live cells require expression of genetically encoded proteins (e.g. FRET sensors). In this work, we describe the development and preliminary application of a set of cell-penetrating, fluorophore labelled peptide substrates for fluorescence lifetime imaging (FLIM) of Abl and Src-family kinase activities. These probes do not rely on FRET pairs or genetically-encoded protein expression. We further demonstrate probe multiplexing and pixel ...[more]

PMID: 33035286

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