Mitochondria Alkylation and Cellular Trafficking Mapped with a Lipophilic BODIPY–Acrolein Fluorogenic Probe
- 13 October 2017
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 139 (45), 16273-16281
- https://doi.org/10.1021/jacs.7b08615
Abstract
Protein and DNA alkylation by endogenously produced electrophiles is associated to the pathogenesis of neurodegenerative diseases, to epigenetic alterations and to cell signaling and redox regulation. With the goal of visualizing, in real-time, the spatiotemporal response of the cell milieu to electrophiles, we have designed a fluorogenic BODIPY-acrolein probe, AcroB, that undergoes a >350-fold fluorescence intensity enhancement concomitant with protein adduct formation. AcroB enables a direct quantification of single post-translational modifications occurring on cellular proteins via recording fluorescence bursts in live-cell imaging studies. In combination with super-resolution imaging, protein alkylation events may be registered and individually counted, yielding a map of protein-electrophile reactions within the cell lipid milieu. Alkylation is predominantly observed within mitochondria, a source, yet not a sink, of AcroB-adducts, illustrating that a mitochondrial constitutive excretion mechanism ensures rapid disposal of compromised proteins. Sorting within the Golgi apparatus and trafficking along microtubules and through the cell endo- and exocytic pathways can be next visualized via live-cell imaging. Our results offer a direct visualization of cellular response to a non-canonical acrolein warhead. We envision AcroB will enable new approaches for diagnosis of pathologies associated to defective cellular trafficking. AcroB may help elucidate key aspects of mitochondria electrophile-adduct excretion, and cell endocytic and exocytic pathways. Conceptually, AcroB provides a new paradigm on fluorescence microscopy studies where chemical perturbation is achieved and simultaneously reported by the probe.Keywords
Funding Information
- Canada Foundation for Innovation
- Natural Sciences and Engineering Research Council of Canada
This publication has 38 references indexed in Scilit:
- Protein Modification by Endogenously Generated Lipid Electrophiles: Mitochondria as the Source and TargetACS Chemical Biology, 2017
- Free Radicals in Biology and MedicinePublished by Oxford University Press (OUP) ,2015
- A Generalizable Platform for Interrogating Target- and Signal-Specific Consequences of Electrophilic Modifications in Redox-Dependent Cell SignalingJournal of the American Chemical Society, 2015
- Quantitative Chemoproteomics for Site-Specific Analysis of Protein Alkylation by 4-Hydroxy-2-Nonenal in CellsAnalytical Chemistry, 2015
- Stable Histone Adduction by 4-Oxo-2-nonenal: A Potential Link between Oxidative Stress and EpigeneticsJournal of the American Chemical Society, 2014
- 4-Hydroxy-2-Nonenal, a Reactive Product of Lipid Peroxidation, and Neurodegenerative Diseases: A Toxic Combination Illuminated by Redox Proteomics StudiesAntioxidants and Redox Signaling, 2012
- Abbott boosts investment in NRF2 activators for reducing oxidative stressNature Reviews Drug Discovery, 2012
- Free Radical Lipid Peroxidation: Mechanisms and AnalysisChemical Reviews, 2011
- Lipid Peroxidation Modification of Protein Generates Nϵ-(4-Oxononanoyl)lysine as a Pro-inflammatory LigandPublished by Elsevier BV ,2011
- Transduction of Redox Signaling by Electrophile-Protein ReactionsScience Signaling, 2009