Geldanamycin Leads to Superoxide Formation by Enzymatic and Non-enzymatic Redox Cycling

The ansamycin antibiotic geldanamycin has frequently been used as an inhibitor of heat shock protein 90 (Hsp90), and this agent has been widely employed as a probe to examine the interactions of Hsp90 with endothelial nitric-oxide synthase. Geldanamycin contains a quinone group, which may participate in redox cycling. When geldanamycin was exposed to the flavin-containing enzyme cytochrome P-450 reductase, both semiquinone and superoxide (O 2 ⨪ ) radicals were detected using electron spin resonance. The treatment of endothelial cells with geldanamycin resulted in a dramatic increase in O 2 ⨪ generation, which was independent of endothelial nitric-oxide synthase, because it was not inhibited byN-nitro-l-arginine methyl ester and also occurred in vascular smooth muscle cells. Diphenylene iodinium inhibited this increase in O 2 ⨪ by 50%, suggesting that flavin-containing enzymes are involved in geldanamycin-induced O 2 ⨪ generation. In the absence of cells, geldanamycin directly oxidized ascorbate, consumed oxygen, and produced O 2 ⨪ . Geldanamycin decreased the bioavailable nitric oxide generated by 3,4-dihydrodiazete 1,2-dioxide in smooth muscle cells by 50%, whereas pretreatment with superoxide dismutase inhibited the effect of geldanamycin. These findings demonstrate that geldanamycin generates O 2 ⨪ , which scavenges nitric oxide, leading to loss of its bioavailability. This effect is independent of the inhibition of Hsp90 and indicates that geldanamycin cannot be used as a specific inhibitor of Hsp90. In light of these findings, the studies using geldanamycin as an inhibitor of Hsp90 should be interpreted with caution.

• Publication year

  2002

• Venue

  Journal of Biological Chemistry

• Publication date

  2002-07-12

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1074/JBC.M203271200PMID 11983711
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• HSP90 as a new therapeutic target for cancer therapy: the story unfolds

  2002cited by this paper
• Hsp90: Chaperoning signal transduction

  2001cited by this paper
• Some Properties of Caldesmon and Calponin and the Participation of These Proteins in Regulation of Smooth Muscle Contraction and Cytoskeleton Formation

  2001cited by this paper
• Heat Shock Protein 90 Mediates the Balance of Nitric Oxide and Superoxide Anion from Endothelial Nitric-oxide Synthase*

  2001cited by this paper
• Hsp90 Ensures the Transition from the Early Ca2+-dependent to the Late Phosphorylation-dependent Activation of the Endothelial Nitric-oxide Synthase in Vascular Endothelial Growth Factor-exposed Endothelial Cells*

  2001cited by this paper
• Estrogen Stimulates Heat Shock Protein 90 Binding to Endothelial Nitric Oxide Synthase in Human Vascular Endothelial Cells

  2000cited by this paper
• Geldanamycin, an inhibitor of heat shock protein 90 (Hsp90) mediated signal transduction has anti‐inflammatory effects and interacts with glucocorticoid receptor in vivo

  2000cited by this paper
• Spin trapping of vascular nitric oxide using colloid Fe(II)-diethyldithiocarbamate.

  2000cited by this paper
• Role of heat shock protein 90 in bradykinin-stimulated endothelial nitric oxide release.

  2000cited by this paper
• Functional Interdependence and Colocalization of Endothelial Nitric Oxide Synthase and Heat Shock Protein 90 in Cerebral Arteries

  2000cited by this paper
• Neuronal Nitric-oxide Synthase Is Regulated by the hsp90-based Chaperone System in Vivo *

  1999cited by this paper
• Hsp90 regulation of endothelial nitric oxide synthase contributes to vascular control in portal hypertension.

  1999cited by this paper
• Hsp90 regulation of endothelial nitric oxide synthase contributes to vascular control in portal hypertension.

  1999cited by this paper
• Superoxide Generation from Endothelial Nitric-oxide Synthase

  1998cited by this paper
• Dynamic activation of endothelial nitric oxide synthase by Hsp90

  1998cited by this paper
• Superoxide production in vascular smooth muscle contributes to oxidative stress and impaired relaxation in atherosclerosis.

  1998cited by this paper
• Oxygen radical generation and enzymatic properties of mitochondria in hypoxia/reoxygenation.

  1998cited by this paper
• Superoxide generation by endothelial nitric oxide synthase: the influence of cofactors.

  1998cited by this paper
• Intracellular hydrogen peroxide and superoxide anion detection in endothelial cells

  1994cited by this paper
• Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycins: essential role for stress proteins in oncogenic transformation.

  1994cited by this paper
• Simulation of multiple isotropic spin-trap EPR spectra.

  1994cited by this paper
• Free radical formation by ansamycin benzoquinone in human breast tumor cells: implications for cytotoxicity and resistance.

  1994cited by this paper
• Role of Iron Ion Chelation by Quinones in Their Reduction, OH-Radical Generation, and Lipid Peroxidation

  1993cited by this paper
• Role of iron ion chelation by quinones in their reduction, OH-radical generation and lipid peroxidation.

  1993cited by this paper
• The reaction of no with superoxide.

  1993cited by this paper
• Role of quinone-iron(III) interaction in NADPH-dependent enzymatic generation of hydroxyl radicals.

  1992cited by this paper
• Vascular bound recombinant extracellular superoxide dismutase type C protects against the detrimental effects of superoxide radicals on endothelium-dependent arterial relaxation.

  1992cited by this paper
• Flow Cytometric Analysis of Respiratory Burst Activity in Phagocytes With Hydroethidine and 2′,7′‐Dichlorofluorescin

  1990cited by this paper
• Free radicals induced by adriamycin-sensitive and adriamycin-resistant cells: a spin-trapping study.

  1989cited by this paper
• Pharmacological evidence that endothelium-derived relaxing factor is nitric oxide: use of pyrogallol and superoxide dismutase to study endothelium-dependent and nitric oxide-elicited vascular smooth muscle relaxation.

  1988cited by this paper
• Streptonigrin-induced deoxyribose degradation: inhibition by superoxide dismutase, hydroxyl radical scavengers and iron chelators.

  1984cited by this paper
• A general mechanism for microsomal activation of quinone anticancer agents to free radicals.

  1978cited by this paper

• The HSP90 inhibitor 17‐AAG induced calcium‐mediated apoptosis in filarial parasites

  2022influential citation
• Amperometric flow-injection analysis of phenols induced by reactive oxygen species generated under daylight irradiation of titania impregnated with horseradish peroxidase.

  2020cites this paper
• Natural heat shock protein 90 inhibitors in cancer and inflammation.

  2020cites this paper
• Mitigating effects of apigenin on edifenphos-induced oxidative stress, DNA damage and apoptotic cell death in human peripheral blood lymphocytes.

  2019cites this paper
• Synergistic effects of geldanamycin with fluconazole are associated with reactive oxygen species in Candida tropicalis resistant to azoles and amphotericin B

  2019cites this paper
• Therapeutic Implications of Heat Shock Proteins in Cancer

  2019cites this paper
• Interaction of C20-substituted derivative of pregnenolone acetate with copper (II) leads to ROS generation, DNA cleavage and apoptosis in cervical cancer cells: Therapeutic potential of copper chelation for cancer treatment.

  2019cites this paper
• Combination of Anti-Cancer Drugs with Molecular Chaperone Inhibitors

  2019cites this paper
• Revisiting silibinin as a novobiocin-like Hsp90 C-terminal inhibitor: Computational modeling and experimental validation.

  2019cites this paper
• Copper-redox cycling by coumarin-di(2-picolyl)amine hybrid molecule leads to ROS-mediated DNA damage and apoptosis: A mechanism for cancer chemoprevention.

  2018cites this paper
• Deciphering the molecular mechanism underlying anticancer activity of coumestrol in triple-negative breast cancer cells.

  2018cites this paper
• Methods to validate Hsp90 inhibitor specificity, to identify off-target effects, and to rethink approaches for further clinical development

  2018cites this paper
• Cytotoxic activity of soy phytoestrogen coumestrol against human breast cancer MCF-7 cells: Insights into the molecular mechanism.

  2017cites this paper
• HSP90 inhibitor 17-DMAG exerts anticancer effects against gastric cancer cells principally by altering oxidant-antioxidant balance

  2017cites this paper
• Cu(II)-coumestrol interaction leads to ROS-mediated DNA damage and cell death: a putative mechanism for anticancer activity.

  2016cites this paper
• Long-term administration of the mitochondria-targeted antioxidant mitoquinone mesylate fails to attenuate age-related oxidative damage or rescue the loss of muscle mass and function associated with aging of skeletal muscle

  2016cites this paper
• A novel quinone derived from 5-hydroxyindoleacetic acid reacts with protein: Possible participation of oxidation of serotonin and its metabolite in the development of atherosclerosis.

  2016cites this paper
• HSP90 Inhibitor Geldanamycin as a Radiation Response Modificator in Human Blood Cells

  2015cites this paper
• The Role of Natural Products in Drug Discovery

  2015cites this paper
• BC265: A Nonquinone Ansamycin Hsp90 Inhibitor Developed Using Biosynthetic Medicinal Chemistry

  2014cites this paper
• N-acetylcysteine prevents the geldanamycin cytotoxicity by forming geldanamycin-N-acetylcysteine adduct.

  2014cites this paper
• Thioredoxin-1 redox signaling regulates cell survival in response to hyperoxia.

  2014cites this paper
• Molecular mechanisms of endothelial NO synthase uncoupling.

  2014cites this paper
• Ascorbic Acid Inhibition of Candida albicans Hsp90-Mediated Morphogenesis Occurs via the Transcriptional Regulator Upc2

  2014cites this paper
• High-Throughput Screen of Natural Product Libraries for Hsp90 Inhibitors

  2014cites this paper
• Synthesis of benzoquinone ansamycin-inspired macrocyclic lactams from shikimic acid.

  2013cites this paper
• Parthenolide generates reactive oxygen species and autophagy in MDA-MB231 cells. A soluble parthenolide analogue inhibits tumour growth and metastasis in a xenograft model of breast cancer

  2013cites this paper
• Cardiomyocyte ryanodine receptor degradation by chaperone-mediated autophagy.

  2013cites this paper
• Natural Product Scaffolds in Cancer Therapy

  2013cites this paper
• Involvement of Calcium-Mediated Reactive Oxygen Species in Inductive GRP78 Expression by Geldanamycin in 9L Rat Brain Tumor Cells

  2013cites this paper
• Hsp90 Is Cleaved by Reactive Oxygen Species at a Highly Conserved N-Terminal Amino Acid Motif

  2012cites this paper
• Redox properties and thiol reactivity of geldanamycin and its analogues in aqueous solutions.

  2012cites this paper
• One-electron reduction of 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin: a pulse radiolysis study.

  2011cites this paper
• Molecular chaperone Hsp90 as a target for oxidant-based anticancer therapies.

  2011influential citation
• Hsp90 is involved in the formation of P-bodies and stress granules.

  2011cites this paper
• EPR detection of cellular and mitochondrial superoxide using cyclic hydroxylamines

  2011cites this paper
• Hsp90 Regulates NADPH Oxidase Activity and Is Necessary for Superoxide but Not Hydrogen Peroxide Production

  2011cites this paper
• Benzoquinone ansamycin 17AAG binds to mitochondrial voltage-dependent anion channel and inhibits cell invasion

  2011cites this paper
• Reactive oxygen species mediate hepatotoxicity induced by the Hsp90 inhibiting anti-cancer geldanamycin and its analogs

  2010influential citation
• Reactive oxygen species mediate hepatotoxicity induced by the Hsp90 inhibitor geldanamycin and its analogs.

  2010cites this paper
• Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin inhibits the proliferation of ARPE-19 cells

  2010cites this paper
• Synthesis of reblastatin, autolytimycin, and non-benzoquinone analogues: potent inhibitors of heat shock protein 90.

  2010cites this paper
• A new feature of the stress response: increase in endocytosis mediated by Hsp70

  2010cites this paper
• Activation of heat shock factor 1 plays a role in pyrrolidine dithiocarbamate-mediated expression of the co-chaperone BAG3.

  2010cites this paper
• 17-allylamino-17-demethoxygeldanamycin inhibits the proliferation of ARPE-19 cells

  2010cites this paper
• Essential Role of Heat Shock Protein 90 in Mediating Non-Genomic Estrogen Signaling in Coronary Artery Smooth Muscle

  2009cites this paper
• Plaque Structure, Prevention and Possible Cure of Alzheimer’s Disease: An Exploratory First Principle Molecular Computational Study

  2009cites this paper
• Hsp90 mediates the balance of nitric oxide and superoxide anion in the lungs of rats with acute pulmonary thromboembolism.

  2009cites this paper
• Essential Role of the 90-Kilodalton Heat Shock Protein in Mediating Nongenomic Estrogen Signaling in Coronary Artery Smooth Muscle

  2009cites this paper
• Role of oxidative stress in geldanamycin-induced cytotoxicity and disruption of Hsp90 signaling complex.

  2009cites this paper
• Natural product inhibitors of Hsp90: potential leads for drug discovery.

  2009cites this paper
• Ansamycin inhibitors of Hsp90: nature's prototype for anti-chaperone therapy.

  2009cites this paper
• Radicicol but not geldanamycin evokes oxidative stress response and efflux protein inhibition in ARPE-19 human retinal pigment epithelial cells.

  2008cites this paper
• Optimizing natural products by biosynthetic engineering: discovery of nonquinone Hsp90 inhibitors.

  2008cites this paper
• Enzymatic Reduction and Glutathione Conjugation of Benzoquinone Ansamycin Heat Shock Protein 90 Inhibitors: Relevance for Toxicity and Mechanism of Action

  2008cites this paper
• Enzymatic reduction and glutathione conjugation of benzoquinone ansamycin Hsp90 inhibitors: Relevance for toxicity and mechanism of action

  2008cites this paper
• Redox Control of Endothelial Function and Dysfunction: Molecular Mechanisms and Therapeutic Opportunities

  2008cites this paper
• Inhibition of brain proinflammatory cytokine synthesis reduces hypothalamic excitation in rats with ischemia-induced heart failure.

  2008cites this paper
• Open Access Research

  2008cites this paper
• Hsp90 inhibitors: Relevance for toxicity and mechanism of action

  2008cites this paper
• One- and Two-Electron-Mediated Reduction of Quinones: Enzymology and Toxicological Implications

  2008cites this paper
• The role of Hsp90 in PC-12 cell survival

  2008cites this paper
• Geldanamycin enhances hepatocyte growth factor stimulation of eNOS phosphorylation in endothelial cells.

  2008cites this paper
• Heat shock protein 90 regulates stabilization rather than activation of soluble guanylate cyclase

  2008cites this paper
• Honokiol is a potent scavenger of superoxide and peroxyl radicals.

  2008cites this paper
• Activation of endothelial nitric oxide synthase by dietary isoflavones: role of NO in Nrf2-mediated antioxidant gene expression.

  2007cites this paper
• Geldanamycin Interferes with the 90-kDa Heat Shock Protein, Affecting Lipopolysaccharide-Mediated Interleukin-1 Expression and Apoptosis within Macrophages

  2007cites this paper
• Electron paramagnetic resonance (EPR) spin trapping of biological nitric oxide.

  2007cites this paper
• Gp91phox-containing NAD(P)H oxidase increases superoxide formation by doxorubicin and NADPH.

  2007cites this paper
• Nitric Oxide and Its Gatekeeper Thrombospondin-1 in Tumor Angiogenesis

  2007cites this paper
• Drug-mediated targeted disruption of multiple protein activities through functional inhibition of the Hsp90 chaperone complex.

  2007cites this paper
• Biotransformation of Geldanamycin and 17-Allylamino-17-Demethoxygeldanamycin by Human Liver Microsomes: Reductive versus Oxidative Metabolism and Implications

  2007cites this paper
• Heat shock protein 90 modulates endothelial nitric oxide synthase activity and vascular reactivity in the newborn piglet pulmonary circulation.

  2007cites this paper
• Production of extracellular superoxide by human lymphoblast cell lines: comparison of electron spin resonance techniques and cytochrome C reduction assay.

  2007cites this paper
• Original Research Communication Mitochondrial Redox Cycling of Mitoquinone Leads to Superoxide Production and Cellular Apoptosis

  2007cites this paper
• Mitochondrial Redox Cycling of Mitoquinone Leads to Superoxide Production and Cellular Apoptosis

  2007cites this paper
• Molecular Mechanisms of Atherosclerosis

  2007influential citation
• Cystine-Glutamate Transporter SLC7A11 Mediates Resistance to Geldanamycin but Not to 17-(Allylamino)-17-demethoxygeldanamycin

  2007cites this paper
• RAR1 and HSP90 Form a Complex with Rac/Rop GTPase and Function in Innate-Immune Responses in Rice[W][OA]

  2007cites this paper
• Geldanamycin , an Inhibitor of Hsp 90 , Potentiates Cytochrome P 4502 E 1-Mediated Toxicity in HepG 2 Cells

  2006cites this paper
• Geldanamycin, an Inhibitor of Hsp90, Potentiates Cytochrome P4502E1-Mediated Toxicity in HepG2 Cells

  2006cites this paper
• Effects of hsp90 binding inhibitors on sGC-mediated vascular relaxation.

  2006cites this paper
• The Bioreduction of a Series of Benzoquinone Ansamycins by NAD(P)H:Quinone Oxidoreductase 1 to More Potent Heat Shock Protein 90 Inhibitors, the Hydroquinone Ansamycins

  2006cites this paper
• Radanamycin, a macrocyclic chimera of radicicol and geldanamycin.

  2006cites this paper
• Hsp90 inhibitors: Small molecules that transform the Hsp90 protein folding machinery into a catalyst for protein degradation

  2006cites this paper
• Increased Superoxide Generation and Decreased Stress Protein Hsp90 Expression in Human Umbilical Cord Vein Endothelial Cells (HUVECs) from Pregnancies Complicated by Preeclampsia

  2006influential citation
• A versatile approach toward the ansamycin antibiotics.

  2006cites this paper
• Geldanamycin, radicicol, and chimeric inhibitors of the Hsp90 N-terminal ATP binding site.

  2006cites this paper
• Interaction of pro‐apoptotic protein HGTD‐P with heat shock protein 90 is required for induction of mitochondrial apoptotic cascades

  2006cites this paper
• Up-regulation of heat shock protein 27 induces resistance to 17-allylamino-demethoxygeldanamycin through a glutathione-mediated mechanism.

  2006cites this paper
• Interactions of peroxynitrite with uric acid in the presence of ascorbate and thiols: implications for uncoupling endothelial nitric oxide synthase.

  2005cites this paper
• Peroxisome proliferator-activated receptor gamma ligands stimulate endothelial nitric oxide production through distinct peroxisome proliferator-activated receptor gamma-dependent mechanisms.

  2005cites this paper
• Endothelial hypoxic preconditioning in rat hypoxic isolated aortic segments

  2005cites this paper
• Roles of 3-Phosphoinositide-dependent Kinase 1 in the Regulation of Endothelial Nitric-oxide Synthase Phosphorylation and Function by Heat Shock Protein 90*

  2005cites this paper
• Chronic ethanol exposure stimulates endothelial cell nitric oxide production through PI-3 kinase-and hsp90-dependent mechanisms.

  2005cites this paper
• Peroxisome Proliferator-Activated Receptor γ Ligands Stimulate Endothelial Nitric Oxide Production Through Distinct Peroxisome Proliferator-Activated Receptor γ–Dependent Mechanisms

  2005cites this paper
• Radester, a novel inhibitor of the Hsp90 protein folding machinery.

  2005cites this paper
• Heat Shock Protein 90 Stabilization of ErbB2 Expression Is Disrupted by ATP Depletion in Myocytes*

  2005cites this paper
• Up‐regulation of the association between heat shock protein 90 and endothelial nitric oxide synthase prevents high glucose‐induced apoptosis in human endothelial cells

  2005cites this paper
• Geldanamycin derivative inhibition of HGF/SF-mediated Met tyrosine kinase receptor-dependent urokinase-plasminogen activation.

  2005cites this paper