36
IRUS TotalDownloads
Altmetric
A coumarin-porphyrin FRET break-apart probe for heme oxygenase-1
File | Description | Size | Format | |
---|---|---|---|---|
jacs.0c12864.pdf | Published version | 2.17 MB | Adobe PDF | View/Open |
SI (FINAL) ja-2020-128646.docx | Supporting information | 6.17 MB | Microsoft Word | View/Open |
Title: | A coumarin-porphyrin FRET break-apart probe for heme oxygenase-1 |
Authors: | Walter, E Ge, Y Mason, J Boyle, J Long, N |
Item 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. |
Issue Date: | 5-May-2021 |
Date of Acceptance: | 30-Mar-2021 |
URI: | http://hdl.handle.net/10044/1/89198 |
DOI: | 10.1021/jacs.0c12864 |
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/) |
Sponsor/Funder: | British Heart Foundation British Heart Foundation British Heart Foundation British Heart Foundation |
Funder's Grant Number: | PG/17/71/33242 FS/13/12/30037 SCRF03 PG/21/10422 |
Keywords: | 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 |
Online Publication Date: | 2021-04-12 |
Appears in Collections: | Chemistry Catalysis and Advanced Materials National Heart and Lung Institute Faculty of Natural Sciences |
This item is licensed under a Creative Commons License