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

Sampreeti Jena; Nur P. Damayanti; Jackie Tan; Erica D. Pratt; Joseph M.K. Irudayaraj; Laurie L. Parker

doi:10.1039/d0cc05030j

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

Sampreeti Jena, Nur P. Damayanti, Jackie Tan, Erica D. Pratt, Joseph M.K. Irudayaraj, Laurie L. Parker

Research output: Contribution to journal › Article › peer-review

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. This journal is

Original language	English (US)
Pages (from-to)	13409-13412
Number of pages	4
Journal	Chemical Communications
Volume	56
Issue number	87
DOIs	https://doi.org/10.1039/d0cc05030j
State	Published - Nov 11 2020

Bibliographical note

Funding Information:
Cells used in this work were a kind gift from Prof. Robert Geahlen (Purdue University Department of Medicinal Chemistry and Molecular Pharmacology). This work was supported by the Purdue Center for Cancer Research (NPD and JI) and the NCI/NIH (R01CA182543 and R33CA217780 to LLP).

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title = "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. This journal is ",

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note = "Funding Information: Cells used in this work were a kind gift from Prof. Robert Geahlen (Purdue University Department of Medicinal Chemistry and Molecular Pharmacology). This work was supported by the Purdue Center for Cancer Research (NPD and JI) and the NCI/NIH (R01CA182543 and R33CA217780 to LLP).",

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AU - Damayanti, Nur P.

AU - Tan, Jackie

AU - Pratt, Erica D.

AU - Irudayaraj, Joseph M.K.

AU - Parker, Laurie L.

N1 - Funding Information: Cells used in this work were a kind gift from Prof. Robert Geahlen (Purdue University Department of Medicinal Chemistry and Molecular Pharmacology). This work was supported by the Purdue Center for Cancer Research (NPD and JI) and the NCI/NIH (R01CA182543 and R33CA217780 to LLP).

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