A Coumarin–Porphyrin FRET Break-Apart Probe for Heme Oxygenase-1 | Zendy

Edward R. H. Walter | Zendy; Ying Ge | Zendy; Justin C. Mason | Zendy; Joseph J. Boyle | Zendy; Nicholas J. Long | Zendy

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A Coumarin–Porphyrin FRET Break-Apart Probe for Heme Oxygenase-1

Author(s) -

Edward R. H. Walter,

Ying Ge,

Justin C. Mason,

Joseph J. Boyle,

Nicholas J. Long

Publication year - 2021

Publication title -

journal of the american chemical society

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 7.115

H-Index - 612

eISSN - 1520-5126

pISSN - 0002-7863

DOI - 10.1021/jacs.0c12864

Subject(s) - chemistry , porphyrin , heme , fluorophore , heme oxygenase , coumarin , förster resonance energy transfer , biophysics , enzyme , fluorescence , hemeprotein , biochemistry , photochemistry , organic chemistry , physics , quantum mechanics , biology

Heme oxygenase-1 (HO-1) is a vital enzyme in humans that primarily regulates free heme concentrations. The overexpression of HO-1 is commonly associated with cardiovascular and neurodegenerative diseases including atherosclerosis and ischemic stroke. Currently, there are no known chemical probes to detect HO-1 activity, limiting its potential as an early diagnostic/prognostic marker in these serious diseases. Reported here are the design, synthesis, and photophysical and biological characterization of a coumarin-porphyrin FRET break-apart probe to detect HO-1 activity, Fe-L 1 . We designed Fe-L 1 to "break-apart" upon HO-1-catalyzed porphyrin degradation, perturbing the efficient FRET mechanism from a coumarin donor to a porphyrin acceptor fluorophore. Analysis of HO-1 activity using Escherichia coli lysates overexpressing hHO-1 found that a 6-fold increase in emission intensity at 383 nm was observed following incubation with NADPH. The identities of the degradation products following catabolism were confirmed by MALDI-MS and LC-MS, showing that porphyrin catabolism was regioselective at the α-position. Finally, through the analysis of Fe-L 2 , we have shown that close structural analogues of heme are required to maintain HO-1 activity. It is anticipated that this work will act as a foundation to design and develop new probes for HO-1 activity in the future, moving toward applications of live fluorescent imaging.

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