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Selective Activation of N , N ′‐Diacyl Rhodamine Pro‐fluorophores Paired with Releasing Enzyme, Porcine Liver Esterase (PLE)
Author(s) -
Abney Kristopher K.,
RamosHunter Susan J.,
Romaine Ian M.,
Goodwin J. Shawn,
Sulikowski Gary A.,
Weaver C. David
Publication year - 2018
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201801409
Subject(s) - fluorophore , rhodamine , fluorescence , esterase , chemistry , rhodamine b , biophysics , enzyme , biochemistry , biology , quantum mechanics , photocatalysis , catalysis , physics
This study reports the synthesis and testing of a family of rhodamine pro‐fluorophores and an enzyme capable of converting pro‐fluorophores to Rhodamine 110. We prepared a library of simple N , N ′‐diacyl rhodamines and investigated porcine liver esterase (PLE) as an enzyme to activate rhodamine‐based pro‐fluorophores. A PLE‐expressing cell line generated an increase in fluorescence rapidly upon pro‐fluorophore addition demonstrating the rhodamine pro‐fluorophores are readily taken up and fluorescent upon PLE‐mediated release. Rhodamine pro‐fluorophore amides trifluoroacetamide (TFAm) and proponamide (PAm) appeared to be the best substrates using a cell‐based assay using PLE expressing HEK293. Our pro‐fluorophore series showed diffusion into live cells and resisted endogenous hydrolysis. The use of our engineered cell line containing the exogenous enzyme PLE demonstrated the rigorousness of amide masking when compared to cells not containing PLE. This simple and selective pro‐fluorophore rhodamine pair with PLE offers the potential to be used in vitro and in vivo fluorescence based assays.