A Targetable Fluorescent Sensor Reveals That Copper-Deficient SCO1 and SCO2 Patient Cells Prioritize Mitochondrial Copper Homeostasis
Top Cited Papers
- 12 May 2011
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 133 (22), 8606-8616
- https://doi.org/10.1021/ja2004158
Abstract
We present the design, synthesis, spectroscopy, and biological applications of Mitochondrial Coppersensor-1 (Mito-CS1), a new type of targetable fluorescent sensor for imaging exchangeable mitochondrial copper pools in living cells. Mito-CS1 is a bifunctional reporter that combines a Cu+-responsive fluorescent platform with a mitochondrial-targeting triphenylphosphonium moiety for localizing the probe to this organelle. Molecular imaging with Mito-CS1 establishes that this new chemical tool can detect changes in labile mitochondrial Cu+ in a model HEK 293T cell line as well as in human fibroblasts. Moreover, we utilized Mito-CS1 in a combined imaging and biochemical study in fibroblasts derived from patients with mutations in the two synthesis of cytochrome c oxidase 1 and 2 proteins (SCO1 and SCO2), each of which is required for assembly and metalation of functionally active cytochrome c oxidase (COX). Interestingly, we observe that although defects in these mitochondrial metallochaperones lead to a global copper deficiency at the whole cell level, total copper and exchangeable mitochondrial Cu+ pools in SCO1 and SCO2 patient fibroblasts are largely unaltered relative to wild-type controls. Our findings reveal that the cell maintains copper homeostasis in mitochondria even in situations of copper deficiency and mitochondrial metallochaperone malfunction, illustrating the importance of regulating copper stores in this energy-producing organelle.Keywords
This publication has 98 references indexed in Scilit:
- Visualizing Ascorbate-Triggered Release of Labile Copper within Living Cells using a Ratiometric Fluorescent SensorJournal of the American Chemical Society, 2010
- Mitochondrial-targeted fluorescent probes for reactive oxygen speciesCurrent Opinion in Chemical Biology, 2009
- Copper redistribution in Atox1-deficient mouse fibroblast cellsJBIC Journal of Biological Inorganic Chemistry, 2009
- Coordination Chemistry of Bacterial Metal Transport and SensingChemical Reviews, 2009
- Application of Metal Coordination Chemistry To Explore and Manipulate Cell BiologyChemical Reviews, 2009
- Structural Biology of Copper TraffickingChemical Reviews, 2009
- A Targetable Fluorescent Probe for Imaging Hydrogen Peroxide in the Mitochondria of Living CellsJournal of the American Chemical Society, 2008
- Mitochondrial copper metabolism and delivery to cytochrome c oxidaseIUBMB Life, 2008
- Molecular Mechanisms of Copper HomeostasisBiochemical and Biophysical Research Communications, 1999
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976