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Biochim Biophys Acta. 2015 Oct;1847(10):1297-309. doi: 10.1016/j.bbabio.2015.07.004. Epub 2015 Jul 14.

Carbon monoxide released by CORM-401 uncouples mitochondrial respiration and inhibits glycolysis in endothelial cells: A role for mitoBKCa channels.

Author information

1
Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Krakow 30-348, Poland. Electronic address: patrycja.kaczara@jcet.eu.
2
INSERM U955, Equipe 12, Créteil, 94000, France; University Paris-Est, Faculty of Medicine, Créteil, 94000, France. Electronic address: roberto.motterlini@inserm.fr.
3
Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD 21201, USA. Electronic address: grosen@umaryland.edu.
4
Laboratory of Intracellular Ion Channels, Nencki Institute of Experimental Biology, Warsaw 02-093, Poland. Electronic address: b.augustynek@nencki.gov.pl.
5
Department of Biophysics, Warsaw University of Life Sciences - SGGW, Warsaw 02-776, Poland. Electronic address: p.bednarczyk@nencki.gov.pl.
6
Laboratory of Intracellular Ion Channels, Nencki Institute of Experimental Biology, Warsaw 02-093, Poland. Electronic address: a.szewczyk@nencki.gov.pl.
7
INSERM U955, Equipe 12, Créteil, 94000, France; University Paris-Est, Faculty of Medicine, Créteil, 94000, France. Electronic address: roberta.foresti@inserm.fr.
8
Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Krakow 30-348, Poland. Electronic address: stefan.chlopicki@jcet.eu.

Abstract

Carbon monoxide (CO), a product of heme degradation by heme oxygenases, plays an important role in vascular homeostasis. Recent evidence indicates that mitochondria are among a number of molecular targets that mediate the cellular actions of CO. In the present study we characterized the effects of CO released from CORM-401 on mitochondrial respiration and glycolysis in intact human endothelial cells using electron paramagnetic resonance (EPR) oximetry and the Seahorse XF technology. We found that CORM-401 (10-100μM) induced a persistent increase in the oxygen consumption rate (OCR) that was accompanied by inhibition of glycolysis (extracellular acidification rate, ECAR) and a decrease in ATP-turnover. Furthermore, CORM-401 increased proton leak, diminished mitochondrial reserve capacity and enhanced non-mitochondrial respiration. Inactive CORM-401 (iCORM-401) neither induced mitochondrial uncoupling nor inhibited glycolysis, supporting a direct role of CO in the endothelial metabolic response induced by CORM-401. Interestingly, blockade of mitochondrial large-conductance calcium-regulated potassium ion channels (mitoBKCa) with paxilline abolished the increase in OCR promoted by CORM-401 without affecting ECAR; patch-clamp experiments confirmed that CO derived from CORM-401 activated mitoBKCa channels present in mitochondria. Conversely, stabilization of glycolysis by MG132 prevented CORM-401-mediated decrease in ECAR but did not modify the OCR response. In summary, we demonstrated in intact endothelial cells that CO induces a two-component metabolic response: uncoupling of mitochondrial respiration dependent on the activation of mitoBKCa channels and inhibition of glycolysis independent of mitoBKCa channels.

KEYWORDS:

CO-RM; Carbon monoxide; Endothelium; Glycolysis; Mitochondrial BKCa channels; Oxidative phosphorylation; Respiration

PMID:
26185029
DOI:
10.1016/j.bbabio.2015.07.004
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