A coumarin-porphyrin FRET break-apart probe for heme oxygenase-1
File(s)
Supporting information
Published version
Author(s)
Walter, Edward
Ge, Ying
Mason, justin
Boyle, Joseph
Long, Nicholas
Type
Journal Article
Abstract
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–L1. We designed Fe–L1 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–L2, 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.
Date Issued
2021-05-05
Date Acceptance
2021-03-30
Citation
Journal of the American Chemical Society, 2021, 143 (17), pp.6460-6469
ISSN
0002-7863
Publisher
American Chemical Society
Start Page
6460
End Page
6469
Journal / Book Title
Journal of the American Chemical Society
Volume
143
Issue
17
Copyright Statement
© 2021 The Authors. Published by American Chemical Society. This is an open access article under a CC-BY Attribution Licence (https://creativecommons.org/licenses/by/4.0/)
License URL
Sponsor
British Heart Foundation
British Heart Foundation
British Heart Foundation
British Heart Foundation
Grant Number
PG/17/71/33242
FS/13/12/30037
SCRF03
PG/21/10422
Subjects
Science & Technology
Physical Sciences
Chemistry, Multidisciplinary
Chemistry
PROTOPORPHYRIN-IX
FLUORESCENT-PROBE
OXIDATION
CANCER
EXPRESSION
COMPLEXES
MECHANISM
MONOXIDE
STROKE
03 Chemical Sciences
General Chemistry
Publication Status
Published
Date Publish Online
2021-04-12