Functional Analysis of Hsp70 Inhibitors

The molecular chaperones of the Hsp70 family have been recognized as targets for anti-cancer therapy. Since several paralogs of Hsp70 proteins exist in cytosol, endoplasmic reticulum and mitochondria, we investigated which isoform needs to be down-regulated for reducing viability of cancer cells. For two recently identified small molecule inhibitors, VER-155008 and 2-phenylethynesulfonamide (PES), which are proposed to target different sites in Hsp70s, we analyzed the molecular mode of action in vitro. We found that for significant reduction of viability of cancer cells simultaneous knockdown of heat-inducible Hsp70 (HSPA1) and constitutive Hsc70 (HSPA8) is necessary. The compound VER-155008, which binds to the nucleotide binding site of Hsp70, arrests the nucleotide binding domain (NBD) in a half-open conformation and thereby acts as ATP-competitive inhibitor that prevents allosteric control between NBD and substrate binding domain (SBD). Compound PES interacts with the SBD of Hsp70 in an unspecific, detergent-like fashion, under the conditions tested. None of the two inhibitors investigated was isoform-specific.

• Publication year

  2013

• Venue

  PLoS ONE

• Publication date

  2013-11-12

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1371/journal.pone.0078443PMID 24265689PMCID 3827032
• 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.

• Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

  2014cited by this paper
• A Modified HSP70 Inhibitor Shows Broad Activity as an Anticancer Agent

  2013cited by this paper
• Allosteric opening of the polypeptide-binding site when an Hsp70 binds ATP

  2013cited by this paper
• A global view of Hsp90 functions

  2013cited by this paper
• Allostery in the Hsp70 chaperone proteins.

  2013influential reference
• Structure and dynamics of the ATP-bound open conformation of Hsp70 chaperones.

  2012cited by this paper
• An interdomain energetic tug-of-war creates the allosterically active state in Hsp70 molecular chaperones.

  2012cited by this paper
• A dancer caught midstep: the structure of ATP-bound Hsp70.

  2012cited by this paper
• Mechanics of Hsp70 chaperones enables differential interaction with client proteins

  2011cited by this paper
• Regulation of HSF1 function in the heat stress response: implications in aging and disease.

  2011cited by this paper
• Overview of the CCP4 suite and current developments

  2011cited by this paper
• Molecular chaperones in protein folding and proteostasis

  2011cited by this paper
• HSP70 Inhibition by the Small-Molecule 2-Phenylethynesulfonamide Impairs Protein Clearance Pathways in Tumor Cells

  2011influential reference
• Differential expression of HSPA1 and HSPA2 proteins in human tissues; tissue microarray-based immunohistochemical study

  2011cited by this paper
• Targeting HSP70: The second potentially druggable heat shock protein and molecular chaperone?

  2010cited by this paper
• The HSP70 chaperone machinery: J proteins as drivers of functional specificity

  2010influential reference
• A novel, small molecule inhibitor of Hsc70/Hsp70 potentiates Hsp90 inhibitor induced apoptosis in HCT116 colon carcinoma cells

  2010cited by this paper
• Identifying and Characterizing Binding Sites and Assessing Druggability

  2009cited by this paper
• A small molecule inhibitor of inducible heat shock protein 70.

  2009influential reference
• The role of molecular chaperones in human misfolding diseases

  2009cited by this paper
• Novel adenosine-derived inhibitors of 70 kDa heat shock protein, discovered through structure-based design.

  2009influential reference
• PILATUS: A single photon counting pixel detector for X-ray applications

  2009cited by this paper
• Hsp110 Is a Nucleotide-activated Exchange Factor for Hsp70*

  2008cited by this paper
• Dual targeting of HSC70 and HSP72 inhibits HSP90 function and induces tumor-specific apoptosis.

  2008cited by this paper
• Allosteric Coupling between the Lid and Interdomain Linker in DnaK Revealed by Inhibitor Binding Studies

  2008cited by this paper
• High-throughput screen for small molecules that modulate the ATPase activity of the molecular chaperone DnaK.

  2008cited by this paper
• Protein disaggregation by the AAA+ chaperone ClpB involves partial threading of looped polypeptide segments

  2008cited by this paper
• Convergence of Molecular, Modeling, and Systems Approaches for an Understanding of the Escherichia coli Heat Shock Response

  2008cited by this paper
• Small-molecule inhibitor of p53 binding to mitochondria protects mice from gamma radiation

  2006cited by this paper
• Structure and mechanism of the Hsp90 molecular chaperone machinery.

  2006cited by this paper
• Allosteric regulation of Hsp70 chaperones by a proline switch.

  2006influential reference
• Designer antibacterial peptides kill fluoroquinolone-resistant clinical isolates.

  2005cited by this paper
• Hsp70 chaperones: Cellular functions and molecular mechanism

  2005influential reference
• Members of the heat-shock protein 70 family promote cancer cell growth by distinct mechanisms.

  2005cited by this paper
• Molecular chaperones and the stress of oncogenesis

  2004cited by this paper
• Phase II study of deoxyspergualin in metastatic breast cancer

  2004cited by this paper
• SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins

  2004cited by this paper
• Coot: model-building tools for molecular graphics.

  2004cited by this paper
• Bag-1M Accelerates Nucleotide Release for Human Hsc70 and Hsp70 and Can Act Concentration-dependent as Positive and Negative Cofactor*

  2001cited by this paper
• Structure of a Bag/Hsc70 Complex: Convergent Functional Evolution of Hsp70 Nucleotide Exchange Factors

  2001cited by this paper
• Stress management - heat shock protein-70 and the regulation of apoptosis.

  2001cited by this paper
• Tuning of chaperone activity of Hsp70 proteins by modulation of nucleotide exchange

  2001influential reference
• Identification of an Inhibitor of hsc70-mediated Protein Translocation and ATP Hydrolysis*

  2001cited by this paper
• Selective depletion of heat shock protein 70 (Hsp70) activates a tumor-specific death program that is independent of caspases and bypasses Bcl-2.

  2000cited by this paper
• Multistep mechanism of substrate binding determines chaperone activity of Hsp70

  2000cited by this paper
• Gene expression profiling of human colon cancer cells following inhibition of signal transduction by 17-allylamino-17-demethoxygeldanamycin, an inhibitor of the hsp90 molecular chaperone

  2000cited by this paper
• Interaction between heat shock proteins and antimicrobial peptides.

  2000cited by this paper
• Heat Shock Protein 70 Is Required for the Survival of Cancer Cells

  2000cited by this paper
• Mechanism of regulation of hsp70 chaperones by DnaJ cochaperones.

  1999cited by this paper
• Selectivity of the molecular chaperone-specific immunosuppressive agent 15-deoxyspergualin: modulation of Hsc70 ATPase activity without compromising DnaJ chaperone interactions.

  1999cited by this paper
• Identification of a binding site on Hsc70 for the immunosuppressant 15-deoxyspergualin.

  1998cited by this paper
• Sequence-specific rates of interaction of target peptides with the molecular chaperones DnaK and DnaJ.

  1998cited by this paper
• Structural Analysis of Substrate Binding by the Molecular Chaperone DnaK

  1996cited by this paper
• Lysine 71 of the Chaperone Protein Hsc70 Is Essential for ATP Hydrolysis*

  1996cited by this paper
• The second step of ATP binding to DnaK induces peptide release.

  1996cited by this paper
• Kinetics of molecular chaperone action.

  1994cited by this paper
• Quantitation of the interaction of the immunosuppressant deoxyspergualin and analogs with Hsc70 and Hsp90.

  1994cited by this paper
• The ATP hydrolysis-dependent reaction cycle of the Escherichia coli Hsp70 system DnaK, DnaJ, and GrpE.

  1994cited by this paper
• Preclinical antitumor activity and pharmacological properties of deoxyspergualin.

  1987cited by this paper

• A chaperone-proteasome-based fragmentation machinery is essential for aggrephagy

  2025influential citation
• Bridging structure and selectivity in chaperone-mediated autophagy: towards targeted therapeutics.

  2025cites this paper
• Dynamics of heat shock protein 70 kDa in heat-shocked and hypoxic human endothelial cells

  2025cites this paper
• HSF1 Activation Mechanisms, Disease Roles, and Small Molecule Therapeutics

  2025cites this paper
• Unraveling the Molecular Inhibition and Conformational Changes of Hsp70 and Hsc70 Induced by VER-155008, a Competitive ATPase Inhibitor through Molecular Dynamics Simulations and Principal Component Analysis

  2025cites this paper
• Disarming the Hsp70-Bim Alliance: Small-Molecule and Peptidic Disruptors of a Chaperone-Apoptotic Switch in Cancer.

  2025cites this paper
• HSP70 governs permeability and mechanotransduction in primary human endothelial cells

  2025cites this paper
• In Silico Analysis of Curcumin and Its Analogs MS13 and MS17 Against HSF1 and HSP Family Proteins

  2025cites this paper
• HSPA2 influences the differentiation and production of immunomodulatory mediators in human immortalized epidermal keratinocyte lines

  2025cites this paper
• Oncogenic stress response mechanisms as new therapeutic targets in cancer treatment: A review

  2025cites this paper
• The dual-function of HSP70 in immune response and tumor immunity: from molecular regulation to therapeutic innovations

  2025cites this paper
• Study on the interventional effect and molecular mechanism of HSP72 in regulating oxidative stress by watermelon seed peptide RDPEER.

  2025cites this paper
• Mechanisms and applications of mitochondrial chaperone inhibitors

  2025cites this paper
• L1, a 3,3′-diindolylmethane-derivative, induced ER stress-mediated apoptosis and suppressed growth through the FLI1/AKT pathway in erythroleukemia HEL cells

  2025cites this paper
• Unveiling the potential of HSPA4: a comprehensive pan-cancer analysis of HSPA4 in diagnosis, prognosis, and immunotherapy

  2024cites this paper
• The human testis-enriched HSPA2 interacts with HIF-1α in epidermal keratinocytes, yet HIF-1α stability and HIF-1-dependent gene expression rely on the HSPA (HSP70) activity.

  2024cites this paper
• Transient Receptor Potential Melastatin 7 (TRPM7) Ion Channel Inhibitors: Preliminary SAR and Conformational Studies of Xenicane Diterpenoids from the Hawaiian Soft Coral Sarcothelia edmondsoni.

  2024cites this paper
• Salidroside directly activates HSC70, revealing a new role for HSC70 in BDNF signalling and neurogenesis after cerebral ischemia

  2024influential citation
• Combined In-Solution Fragment Screening and Crystallographic Binding-Mode Analysis with a Two-Domain Hsp70 Construct.

  2024cites this paper
• Examination of Triazolinedione-Derived Dihydropyrrole Hybrid Compounds: ER Stress-Related Apoptosis in Breast Cancer Cells, Molecular Docking, and ADMET Analysis.

  2024cites this paper
• Heat shock cognate 70 protein is a novel target of nobiletin and its colonic metabolites in inhibiting colon carcinogenesis.

  2024cites this paper
• The regulation of the thermal stability and affinity of the HSPA5 (Grp78/BiP) by clients and nucleotides is modulated by domains coupling.

  2024cites this paper
• HSPA8 inhibitors augment cancer chemotherapeutic effectiveness via potentiating necroptosis

  2024influential citation
• The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds in a cellular system

  2023cites this paper
• HSP70 inhibitor amplifies the bFGF‑induced release of IL‑6 in osteoblasts.

  2023cites this paper
• Enhanced Membrane Fluidization and Cholesterol Displacement by 1-Heptanol Inhibit Mast Cell Effector Functions

  2023cites this paper
• The multifunction of HSP70 in cancer: Guardian or traitor to the survival of tumor cells and the next potential therapeutic target.

  2023cites this paper
• A biosensor of protein foldedness identifies increased “holdase” activity of chaperones in the nucleus following increased cytosolic protein aggregation

  2022cites this paper
• The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds

  2022cites this paper
• Allosteric HSP70 inhibitors perturb mitochondrial proteostasis and overcome proteasome inhibitor resistance in multiple myeloma.

  2022cites this paper
• HSP70 inhibitors upregulate prostaglandin E1-induced synthesis of interleukin-6 in osteoblasts

  2022cites this paper
• Discovery and Development of a Selective Inhibitor of the ER Resident Chaperone Grp78.

  2022cites this paper
• Comparative analysis of the anticoagulant activities and immunogenicity of HSC70 and HSC70TKD of Haemaphysalis flava

  2022cites this paper
• Identification of new small molecules as dual FoxM1 and Hsp70 inhibitors using computational methods

  2022cites this paper
• N-myc mediated translation control is a therapeutic vulnerability in medulloblastoma.

  2022cites this paper
• Pharmacological agents selectively acting on the channel moieties of TRPM6 and TRPM7.

  2022cites this paper
• Advances in the study of HSP70 inhibitors to enhance the sensitivity of tumor cells to radiotherapy

  2022cites this paper
• Requirement of Peptidyl-Prolyl Cis/Trans isomerases and chaperones for cellular uptake of bacterial AB-type toxins

  2022cites this paper
• Cellular aging impedes stress-activation of a crucial JDP-Hsp70 protein disaggregase

  2022cites this paper
• CHIP ubiquitin ligase is involved in the nucleolar stress management

  2022cites this paper
• Protein painting reveals pervasive remodeling of conserved proteostasis machinery in response to pharmacological stimuli

  2022cites this paper
• Non-cell-autonomous HSC70.1 chaperone displays homeostatic feed-back regulation by binding its own mRNA

  2022cites this paper
• A Census of Hsp70-Mediated Proteome Solubility Changes upon Recovery from Heat Stress.

  2022cites this paper
• Functional Effects of ARV-1502 Analogs Against Bacterial Hsp70 and Implications for Antimicrobial Activity

  2022cites this paper
• Heat Shock Protein Member 8 (HSPA8) Is Involved in Porcine Reproductive and Respiratory Syndrome Virus Attachment and Internalization

  2022cites this paper
• A biosensor to gauge protein homeostasis resilience differences in the nucleus compared to cytosol of mammalian cells

  2021cites this paper
• Distinguishing the optimal binding mechanism through reversible and irreversible inhibition analysis of HSP72 protein in cancer therapy

  2021cites this paper
• An allosteric inhibitor of bacterial Hsp70 chaperone potentiates antibiotics and mitigates resistance.

  2021cites this paper
• Clinicopathological significance of HSP70 expression in gastric cancer: a systematic review and meta-analysis

  2021cites this paper
• Unsymmetrical Trifluoromethyl Methoxyphenyl β-Diketones: Effect of the Position of Methoxy Group and Coordination at Cu(II) on Biological Activity

  2021cites this paper
• Heat Shock Cognate 70 kDa Protein Is the Target of Tetradecyl 2,3-Dihydroxybenzoate for Neuritogenic Effect in PC12 Cells

  2021cites this paper
• Novel Antibacterial Targets in Protein Biogenesis Pathways

  2021cites this paper
• Heat Shock Protein 70 as a Sex-Skewed Regulator of α-Synucleinopathy

  2021cites this paper
• Arginine-Arginine-Leucine Peptide Targeting Heat Shock Protein 70 for Cancer Imaging.

  2021cites this paper
• Comparison of irreversible inhibition targeting HSP72 protein: the resurgence of covalent drug developments

  2021cites this paper
• Inhibition of the Heat Shock Protein A (HSPA) Family Potentiates the Anticancer Effects of Manumycin A

  2021cites this paper
• Function, Therapeutic Potential, and Inhibition of Hsp70 Chaperones.

  2021cites this paper
• Turkish Computational and Theoretical Chemistry

  2021cites this paper
• Microthermal-induced subcellular-targeted protein damage in cells on plasmonic nanosilver-modified surfaces evokes a two-phase HSP-p97/VCP response

  2021cites this paper
• Cell surface heat shock protein-mediated entry of tumor cell-adapted rotavirus into U-937 cells

  2021cites this paper
• 2-Phenylethyne-1-Sulfonamide Derivatives as New Drugs Candidates for Heat Shock Protein 70 and Doublecortin-like Kinase

  2021cites this paper
• Hsp70 acts as a fine-switch that controls E3 ligase CHIP-mediated TAp63 and ΔNp63 ubiquitination and degradation

  2021cites this paper
• Dissecting the interaction between HSP70 and vascular contraction: role of $$\hbox{Ca}^{2+}$$ Ca 2 + handling mechanisms

  2021cites this paper
• 2-phenylethynesulfonamide inhibits growth of oral squamous cell carcinoma cells by blocking the function of heat shock protein 70

  2020cites this paper
• Widespread remodeling of proteome solubility in response to different protein homeostasis stresses

  2020cites this paper
• The heat shock protein 70 inhibitor VER155008 suppresses the expression of HSP27, HOP and HSP90β and the androgen receptor, induces apoptosis, and attenuates prostate cancer cell growth

  2020cites this paper
• Rapid and simple screening of the apoptotic compounds based on Hsp70 inhibition using luciferase as an intracellular reporter

  2020cites this paper
• Compilation of antiviral treatments and strategies to fight fish viruses

  2020cites this paper
• Role of heat-shock proteins in infection of human adenocarcinoma cell line MCF-7 by tumor-adapted rotavirus isolates

  2020cites this paper
• PES inhibits human-inducible Hsp70 by covalent targeting of cysteine residues in the substrate-binding domain

  2020cites this paper
• Intoxication of mammalian cells with binary clostridial enterotoxins is inhibited by the combination of pharmacological chaperone inhibitors

  2020cites this paper
• Shear stress-exposed pulmonary artery endothelial cells fail to upregulate HSP70 in chronic thromboembolic pulmonary hypertension

  2020cites this paper
• Heat Shock Protein and Cancer Based Therapies

  2020cites this paper
• Hsp70 in cancer: A double agent in the battle between survival and death

  2020cites this paper
• HSPA2 Chaperone Contributes to the Maintenance of Epithelial Phenotype of Human Bronchial Epithelial Cells but Has Non-Essential Role in Supporting Malignant Features of Non-Small Cell Lung Carcinoma, MCF7, and HeLa Cancer Cells

  2020influential citation
• Heat shock protein 70 positively regulates transforming growth factor-α-induced hepatocellular carcinoma cell migration via the AKT signaling pathway

  2020cites this paper
• Coupling of HSP72 α-Helix Subdomains by the Unexpected Irreversible Targeting of Lysine-56 over Cysteine-17; Coevolution of Covalent Bonding

  2020cites this paper
• Structural basis of DegP protease temperature-dependent activation

  2020cites this paper
• S-Glutathionylation of human inducible Hsp70 reveals a regulatory mechanism involving the C-terminal α-helical lid

  2020cites this paper
• S-Glutathionylation of human inducible Hsp70 reveals a regulatory mechanism involving the C-terminal α-helical lid.

  2020cites this paper
• Reflections and Outlook on Targeting HSP90, HSP70 and HSF1 in Cancer: A Personal Perspective.

  2020cites this paper
• HSF1 and Molecular Chaperones in Biology and Cancer

  2020cites this paper
• Necroptosis-blocking compound NBC1 targets heat shock protein 70 to inhibit MLKL polymerization and necroptosis

  2020cites this paper
• Autophagic Inhibition via Lysosomal Integrity Dysfunction Leads to Antitumor Activity in Glioma Treatment

  2020cites this paper
• Lysosomes as a therapeutic target

  2019cites this paper
• Inhibition of Hsp70 Suppresses Neuronal Hyperexcitability and Attenuates Epilepsy by Enhancing A-Type Potassium Current.

  2019cites this paper
• Hsp70 interactions with membrane lipids regulate cellular functions in health and disease.

  2019cites this paper
• Heat shock protein 70 dampens the inflammatory response of human PDL cells to mechanical loading in vitro.

  2019cites this paper
• DnaJB6 is a RanGTP-regulated protein involved in dynein-dependent microtubule organization during mitosis

  2019cites this paper
• Isolation and characterization of a novel scFv antibody fragments specific for Hsp70 as a tumor biomarker

  2019cites this paper
• DnaJB6 is a RanGTP-regulated protein required for microtubule organization during mitosis

  2019cites this paper
• A high throughput substrate binding assay reveals hexachlorophene as an inhibitor of the ER-resident HSP70 chaperone GRP78.

  2019cites this paper
• 2‐ and N6‐functionalized adenosine‐5′‐diphosphate analogs for the inhibition of mortalin

  2019cites this paper
• The Molecular Chaperone Heat Shock Protein 70 Controls Liver Cancer Initiation and Progression by Regulating Adaptive DNA Damage and Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Signaling Pathways

  2019cites this paper
• The nucleolus functions as a phase-separated protein quality control compartment

  2019cites this paper
• Widespread remodelling of proteome solubility in response to different protein homeostasis stresses

  2019cites this paper
• HSPA8/HSC70 in Immune Disorders: A Molecular Rheostat that Adjusts Chaperone-Mediated Autophagy Substrates

  2019cites this paper
• Hsp70 inhibitors: Implications for the treatment of colorectal cancer

  2019influential citation
• Heat Shock Proteins and Ovarian Cancer: Important Roles and Therapeutic Opportunities

  2019cites this paper
• Functional redundancy of HSPA1, HSPA2 and other HSPA proteins in non-small cell lung carcinoma (NSCLC); an implication for NSCLC treatment

  2019cites this paper