Oligomycin A

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Oligomycin A
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Executive Summary Information

Compound Oligomycin A
Toxicities Cytotoxicity
Mechanisms Oligomycin A is a natural product that is a specific inhibitor of complex V (ATP synthase) of the electron transport chain.
Comments This compound is an MOA standard for inhibition of mitochondrial function.
Feedback Contact Gold Compound Working Group (GCWG)
Oligomycin A
Oligomycin.png
Identifiers
Leadscope Id LS-98326
CAS 1404-19-9
ChemSpider 21106358
Pathway DBs
Assay DBs
PubChem CID 6450197
Omics DBs
Properties
ToxCast Accepted no
Toxic Effect Cytotoxicity
ToxBank Accepted yes


In Vivo Data ? Compound Assessment
Adverse Events ? Oligomycin A is generally lethal to all eukaryotes.
Toxicity Mechanisms ? Oligomycin A is a Streptomyces macrolide that binds to the oligomycin sensitivity-conferring protein (OSCP) at the F(o) subunits 6 and 9 which are found in the stalk of the F1F0-ATPase complex. This binding blocks proton conductance across the synthase complex and inhibits the synthesis of mitochondrial ATP. As an additional result, the proton pumps of the electron transport chain are unable to operate because the gradient becomes too strong for them to overcome. NADH is then no longer oxidized and the citric acid cycle ceases to operate because the concentration of NAD+ falls below the concentration that these enzymes can use.

References:

-Joshi S and Huang YG (1991). "ATP synthase complex from bovine heart mitochondria: the oligomycin sensitivity conferring protein is essential for dicyclohexyl carbodiimide-sensitive ATPase". Biochim. Biophys. Acta 1067 (2): 255–258. pmid:1831660 .

Since oligomycin blocks oxidative phosphorylation, it makes cells more dependent on glycolysis and more sensitive to inhibitors of glycolysis such as 2-deoxy-D-glucose. Cells that are deficient in HIF-1 α, which induces increased glucose uptake and glycolysis, are more sensitive to the toxicity of oligomycin whereas cells that are sufficient in HIF-1α are less sensitive to oligomycin under hypoxic conditions (where HIF-1α accumulates). However, because oligomycin blocks reduction of O2 by the electron transport, it maintains higher levels of O2 and modulates down the protective effect of HIF-1α under normoxic conditions.

References:

-Frederic Dayan, Rebecca L. Bilton, Julie Laferriere, Eric Trottier, Daniele Roux, Jacques Pouyssegur, and Nathalie. M. Mazure, “Activation of HIF-1a in Exponentially Growing Cells Via Hypoxic Stimulation Is Independent of the Akt/mTOR Pathway” J. Cell. Physiol. 218: 167–174, 2009.
-Johnathan C. Maher, Medhi Wangpaichitr, Niramol Savaraj, Metin Kurtoglu, and Theodore J. Lampidis, “Hypoxia-inducible factor-1 confers resistance to the glycolytic inhibitor 2-deoxy-D-glucose”, Mol Cancer Ther (2007) 6:732–41.
-Arthur R. Salomon, David W. Voehringer, Leonard A. Herzenberg, and Chaitan Khosla, “Understanding and exploiting the mechanistic basis for selectivity of polyketide inhibitors of F0F1-ATPase”,PNAS (2000) 97:14766–14771.
-Gong Y, Agani FH.,“Oligomycin inhibits HIF-1alpha expression in hypoxic tumor cells”, Am J Physiol Cell Physiol. 2005 May;288(5):C1023-9.

Oligomycin is a key tool for measuring mitochondrial (dys)function, appropriate to assessing the effect of other toxicants on mitochondrial function. In many cultured cells, there is sufficient capacity for glycolysis to maintain viability despite inhibition of oxidative phosphorylation. In these cells, treatment with oligomycin, and subsequently with FCCP, rotenone, mixothiazol, and ionophores, can be used to assess capacity for NADH formation, flux through the electron transport chain, and maintenance of the mitochondrial membrane potential. Brand and Nicholls (2011) provide an excellent review of the effects of oligomycin alone and in conjunction with other inhibitors on mitochondrial function.

References:

-Martin D. Brand and David G. Nicholls, “Assessing mitochondrial dysfunction in cells”, Biochem. J. (2011) 435:297–312.

The ATP synthase reaction is close enough to equilibrium so that under conditions of ischemia, e.g. in cardiomyocytes, the synthase runs in reverse as an ATPase. Uncouplers such as FCCP mimic this effect, so that dispersing the mitochondrial proton gradient not only blocks mitochondrial ATP synthesis but drives the ATP synthase in the direction of ATP hydrolysis to actively deplete ATP, accounting for 50-80% of total ATP utilization. Since oligomycin also blocks the ATPase activity of complex V, it is protective vs. ischemia or FCCP. Because oligomycin protects only against ATP depletion and not depletion of the proton gradient, it is concluded that cytotoxicity of inhibitors of oxidative phosphorylation reflects depletion of ATP and not depletion of the mitochondrial membrane potential.

References:

-Robert B. Jennings, Keith a. Reimer, and Charles Steenbergen, “Effect of Inhibition of the Mitochondrial ATPase on Net Mycocardial ATP in Total Ischemia”, J Mol Cell Cardiol (1991) 23:1383-1395.
-A. L. Nieminen, A. K. Saylor, B. Herman, and J. J. Lemasters, “ATP depletion rather than mitochondrial depolarization mediates hepatocyte killing after metabolic inhibition”, Am J Physiol Cell Physiol July 1, (1994) 267:C67-C74.
-Gary J. Grover, Karnail S. Atwal, Paul G. Sleph, Feng-Li Wang, Hossain Monshizadegan, Thomas Monticello, and David W. Green, “Excessive ATP hydrolysis in ischemic myocardium by mitochondrial F1F0-ATPase: effect of selective pharmacological inhibition of mitochondrial ATPase hydrolase activity”, Am J Physiol Heart Circ Physiol 287:H1747–H1755 (2004).

Oligomycin protects against TNF-induced apoptosis in a manner very similar to protection vs. FCCP toxicity (above) and consistent with increased permeability of the mitochondrial membrane as the mechanism of TNF-induced apoptosis.

References:

-Liarisa A Shchepina, Olga Y Pletjushkina, Armine V Avetisyan, Liora E Bakeeva, Elena K Fetisova, Denis S Izyumov, Valeria B Saprunova, Mikhail Yu Vyssokikh, Boris V Chernyak, and Vladimir P Skulach, “Oligomycin, inhibitor of the F0 part of H+-ATP-synthase, suppresses the TNF-induced apoptosis” Oncogene (2002) 21, 8149 – 8157.
-Kathryn M. Johnson, Lara Swenson, Anthony W. Opipari Jr., Rolf Reuter, Nawid Zarrabi, Carol A. Fierke, Michael Börsch, and Gary D. Glick, “Mechanistic Basis for Differential Inhibition of the F1Fo-ATPase by Aurovertin”, Biopolymers (2009) 91: 830–840.

Oligomycin at high concentrations (IC50 = 4.5 uM) may also inhibit the plasma membrane Na+-K+-ATPase. Although oligomycin stimulates Na+ binding to Na+/K+-ATPase, it inhibits Na+/Na+ exchange, and does not affect either Na+-dependent AD/-ATP exchange or K+-dependent phosphatase activity.

References:

-Arato-Oshima T, Matsui H, Wakizaka A, Homareda H., “Mechanism responsible for oligomycin-induced occlusion of Na+ within Na/K-ATPase” (1996) J. Biol. Chem. 271, 25604.
-Fahn, S, Koval, GJ, and Albers RW, “Sodium-Potassium-activated Adenosine Triphosphatase of Electrophorus Electric Organ”, (1966) J. Biol. Chem. 241, 1882.

Therapeutic Target ? Oligomycin A is used to modulate ATP synthesis in studies of cell or organ function. It is not used therapeutically.



PubMed references

The following resource link will perform a PubMed query for the terms "Oligomycin A" and "mytochondrial function".
FCCP Search

References


PK-ADME ? Compound Assessment
PK parameters ? Data not found. In vivo studies are limited due to poor solubility.
Therapeutic window ?
Mouse LD501.5 mg/kg ip (2 umol/kb)
10 mg/kg po

References:

-Santa Cruz Biotechnology, Inc: MSDS
-Genaxxon Bioscience: MSDS

Metabolically activated ? The parent compound is the toxic species.

Omics and IC50 Data ? Compound Assessment
Gene expression profiles known. ? References:
-Sandra Varum, Ana S. Rodrigues, Michelle B. Moura, Olga Momcilovic, Charles A. Easley IV, João Ramalho-Santos, Bennett Van Houten, Gerald Schatten, “Energy Metabolism in Human Pluripotent Stem Cells and Their Differentiated Counterparts”
-Justin Lamb, Emily D. Crawford, David Peck, Joshua W. Modell, Irene C. Blat, Matthew J. Wrobel, Jim Lerner, Jean-Philippe Brunet, Aravind Subramanian, Kenneth N. Ross, Michael Reich, Haley Hieronymus, Guo Wei1, Scott A. Armstrong, Stephen J. Haggarty, Paul A. Clemons, Ru Wei, Steven A. Carr, Eric S. Lander, and Todd R. Golub, “The Connectivity Map: Using Gene-Expression Signatures to Connect Small Molecules, Genes, and Disease”, Science (2006) 313:1929-1935.
-Gene expression patterns across NCI cell lines: Arthur R. Salomon, David W. Voehringer, Leonard A. Herzenberg, and Chaitan Khosla, “Understanding and exploiting the mechanistic basis for selectivity of polyketide inhibitors of F0F1-ATPase”,PNAS (2000) 97:14766–14771.
-Gong Y, Agani FH.,“Oligomycin inhibits HIF-1alpha expression in hypoxic tumor cells”, Am J Physiol Cell Physiol. 2005 May;288(5):C1023-9.
-Emmanuel Chevillotte, Marta Giralt, Bruno Miroux, Daniel Ricquier, and

Francesc Villarroya, “Uncoupling Protein-2 Controls Adiponectin Gene Expression in Adipose Tissue Through the

Modulation of Reactive Oxygen Species Production”, Diabetes (2007) 56:1042-1050.
-Xiong W, Jiao Y, Huang W, Ma M, Yu M, Cui Q, Tan D., “Regulation of the cell cycle via mitochondrial gene expression and energy metabolism in HeLa cells”, Acta Biochim Biophys Sin (Shanghai). 2012 Apr;44(4):347-58.
-Bergstraesser C, Hoeger S, Song H, Ermantraut L, Hottenrot M, Czymai T, Schmidt M, Goebeler M, Ponelies N, Stich C, Loesel R, Molema G, Seelen M, van Son W, Yard BA, Rafat N., “Inhibition of VCAM-1 expression in endothelial cells by CORM-3: the role of the ubiquitin-proteasome system, p38, and mitochondrial respiration”, Free Radic Biol Med. 2012 Feb 15;52(4):794-802. Epub 2011 Dec 21.
Proteomics profiles known. ? References:
-Frederic Dayan, Rebecca L. Bilton, Julie Laferriere, Eric Trottier, Daniele Roux, Jacques Pouyssegur, and Nathalie. M. Mazure, “Activation of HIF-1a in Exponentially Growing Cells Via Hypoxic Stimulation Is Independent of the Akt/mTOR Pathway”. J. Cell. Physiol. 218: 167–174, 2009.
-Johnathan C. Maher, Medhi Wangpaichitr, Niramol Savaraj, Metin Kurtoglu, and Theodore J. Lampidis, “Hypoxia-inducible factor-1 confers resistance to the glycolytic inhibitor 2-deoxy-D-glucose”, Mol Cancer Ther (2007) 6:732–41.
-Gong Y, Agani FH.,“Oligomycin inhibits HIF-1alpha expression in hypoxic tumor cells”, Am J Physiol Cell Physiol. 2005 May;288(5):C1023-9.
Metabonomics profiles known. ? References:
-Effects on ATP synthesis: Martin D. Brand and David G. Nicholls, “Assessing mitochondrial dysfunction in cells”, Biochem. J. (2011) 435:297–312.
Fluxomics profiles known. ?
Epigenomics profiles known. ?
Observed IC50 for in vitro cellular efficacy. ? Not Applicable
Observed IC50 for in vitro cellular toxicity studies. ? Kd = 1.7 nM for binding to the oligomycin sensitivity-conferring protein (OSCP) from bovine heart.

References:

-Alain Dupuis, Michel Satre, and Pierre V. Vignais, “Titration of the binding sites for the oligomycin-sensitivity conferring protein in beef heart submitochondrial particles”, FEBS Letters (1983) 156:99-102.

IC50 = 8 nM for inhibition of ATP synthase in beef heart mitochondria

References:

-Sashi Nadanaciva, Autumn Bernal, Robert Aggeler, Roderick Capaldi, Yvonne Will, “Target identiWcation of drug induced mitochondrial toxicity using immunocapture based OXPHOS activity assays”, Toxicology in Vitro (2007) 21:902-11

Typical concentrations for inhibition of oxidative phosphorylation in cell culture studies are 1-5 ug/mL (1-5 uM).

References:

-Choi, S. W., Gerencser, A. A. and Nicholls, D. G. (2009) “Bioenergetic analysis of isolated cerebrocortical nerve terminals on a microgram scale: spare respiratory capacity and stochastic mitochondrial failure”, J. Neurochem. 109, 1179–1191.
-Ward, M. W., Rego, A. C., Frenguelli, B. G. and Nicholls, D. G. (2000) “Mitochondrial membrane potential and glutamate excitotoxicity in cultured cerebellar granule cells”, J. Neurosci. 20, 7208–7219
-Frederic Dayan, Rebecca L. Bilton, Julie Laferriere, Eric Trottier, Daniele Roux, Jacques Pouyssegur, and Nathalie. M. Mazure, “Activation of HIF-1a in Exponentially Growing Cells Via Hypoxic Stimulation Is Independent of the Akt/mTOR Pathway” J. Cell. Physiol. 218: 167–174, 2009.

In cells with insufficient capacity for glycolysis to protect against cytotoxicity of oligomycin, the maximum cell death has been observed at 0.1 ug/mL.

References:

-A. L. Nieminen, A. K. Saylor, B. Herman, and J. J. Lemasters, “ATP depletion rather than mitochondrial depolarization mediates hepatocyte killing after metabolic inhibition”, Am J Physiol Cell Physiol July 1, (1994) 267:C67-C74.


IC50 = 4.5 uM for inhibition of the plasma membrane Na+-K+-ATPase.

References:

-Arato-Oshima T, Matsui H, Wakizaka A, Homareda H., “Mechanism responsible for oligomycin-induced occlusion of Na+ within Na/K-ATPase” (1996) J. Biol. Chem. 271, 25604.
-Fahn, S, Koval, GJ, and Albers RW, “Sodium-Potassium-activated Adenosine Triphosphatase of Electrophorus Electric Organ”, (1966) J. Biol. Chem. 241, 1882.

Physical Properties ? Compound Assessment
Accepted and listed within the ToxCast/Tox21 program. ? No - Not included in ToxCast Phase I and II Chemicals List
Substance stability. ? Solid is stable for 2-3 weeks at room temperature, for up to 2 years at -20°C. Conditions to avoid: acid, base, light and heat.

References:

-MSDS Cell Signaling Technology

Oligomycin is fairly stable in solution. It should be stable dissolved in ethanol for at least a week at 0-4 °C and for several months when stored frozen in aliquots at -20 °C.

References:

-Sigma Aldrich (O4876) Product information
-Enzo Lifescience Oligomycin A datasheet

Recommend that aqueous solutions be made fresh daily

References:

-Cayman Chemical product sheet
Soluble in buffer solution at 30 times the in vitro IC50 for toxicity. ? Water: 0.02 – 0.2 mg/mL (25°C) (25 - 250 uM)

References:

-Santa Cruz Biotechnology, Inc MSDS
-Genaxxon Bioscience MSDS


estimated intrinsic solubility : 5 mg/ml
estimated solubility in pure water at pH 6.96: 5 mg/ml
estimated solubility in water at pH 7.4: 5 mg/ml
Calculations performed using ACD/PhysChem v 12.0

Solubility in DMSO 100 times buffer solubility. ? 50 – 300 mg/mL (0.1 – 0.5 M)

References:

-[http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Product_Information_Sheet/2/o4876pis.Par.0001.File.tmp/o4876pis.pdf Sigma Aldrich (O4876) Product information]
-Enzo Lifescience Oligomycin A datasheet
Vessel binding properties. ? Probable
Vapor pressure. (Non-volatile) ? estimated vapor pressure (25°C): 5.74E-031 mmHg (Calculation performed using EPI Suite v4.10)


Authors of this ToxBank wiki page

David Bower, Egon Willighagen
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