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2014 ; 15
(9
): 1317-24
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Fluorescent visualization of Src by using dasatinib-BODIPY
#MMPMID24828915
Vetter ML
; Zhang Z
; Liu S
; Wang J
; Cho H
; Zhang J
; Zhang W
; Gray NS
; Yang PL
Chembiochem
2014[Jun]; 15
(9
): 1317-24
PMID24828915
show ga
Many biological experiments are not compatible with the use of
immunofluorescence, genetically encoded fluorescent tags, or FRET-based
reporters. Conjugation of existing kinase inhibitors to cell-permeable
fluorophores can provide a generalized approach to develop fluorescent probes of
intracellular kinases. Here, we report the development of a small molecule probe
of Src through conjugation of BODIPY to two well-established dual Src-Abl kinase
inhibitors, dasatinib and saracatinib. We show that this approach is not
successful for saracatinib but that dasatinib-BODIPY largely retains the
biological activity of its parent compound and can be used to monitor the
presence of Src kinase in individual cells by flow cytometry. It can also be used
to track the localization of Src by fixed and live-cell fluorescence microscopy.
This strategy could enable generation of additional kinase-specific probes useful
in systems not amenable to genetic manipulation or could be used together with
fluorescent proteins to enable a multiplexed assay readout.
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