A systematic protocol for the characterization of Hsp90 modulators

Matts, Robert L.; Brandt, Gary E. L.; Lu, Yue; Dixit, Anshuman; Mollapour, Mehdi; Wang, Suiquan; Donnelly, Alison C.; Neckers, Leonard Μ.; Verkhivker, Gennady M.; Blagg, Brian S. J.

doi:10.1016/j.bmc.2010.10.029

Cited by 78 publications

(85 citation statements)

References 46 publications

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“…The molecular mechanism of action related to the biologic effects of gedunin relies on the inhibition of Hsp90 activity via specific binding to the cochaperone p23 and by disrupting the cochaperone Cdc37-Hsp90 interaction (Matts et al, 2011;Patwardhan et al, 2013). However, here we demonstrate that gedunin mechanisms of action go beyond Hsp90 modulation.…”

Section: Introductionmentioning

confidence: 56%

“…Hsp90 is an abundantly and ubiquitously expressed chaperone that helps to maintain, at the expense of ATP, the structure of several membrane, cytoplasmic, and endoplasmic reticulumassociated client proteins (Pratt and Toft, 1997;Picard, 2002;Zhao and Houry, 2005;McClellan et al, 2007). Hsp90 chaperoning activity can be inhibited by geldanamycin, 17-allylamino-17-demethoxy-geldanamycin (17-AAG), and celastrol, which leads to increased degradation of Hsp90 client proteins (Matts et al, 2011). In experimental models, Hsp90 inhibitors have been shown to suppress different signaling pathways and display potent antiproliferative, cytoprotective, and anti-inflammatory activities (Lewis et al, 2000;Poulaki et al, 2007;Ambade et al, 2012;Chow et al, 2013;Leung et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Gedunin Binds to Myeloid Differentiation Protein 2 and Impairs Lipopolysaccharide-Induced Toll-Like Receptor 4 Signaling in Macrophages

et al. 2015

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Recognition of bacterial lipopolysaccharide (LPS) by innate immune system is mediated by the cluster of differentiation 14/ Toll-like receptor 4/myeloid differentiation protein 2 (MD-2) complex. In this study, we investigated the modulatory effect of gedunin, a limonoid from species of the Meliaceae family described as a heat shock protein Hsp90 inhibitor, on LPS-induced response in immortalized murine macrophages. The pretreatment of wild-type (WT) macrophages with gedunin (0.01-100 mM, noncytotoxic concentrations) inhibited LPS (50 ng/ml)-induced calcium influx, tumor necrosis factor-a, and nitric oxide production in a concentration-dependent manner. The selective effect of gedunin on MyD88-adapter-like/myeloid differentiation primary response 88-and TRIF-related adaptor molecule/TIR domain-containing adapter-inducing interferon-b-dependent signaling pathways was further investigated. The pretreatment of WT, TIR domain-containing adapter-inducing interferon-b knockout, and MyD88 adapter-like knockout macrophages with gedunin (10 mM) significantly inhibited LPS (50 ng/ml)-induced tumor necrosis factor-a and interleukin-6 production, at 6 hours and 24 hours, suggesting that gedunin modulates a common event between both signaling pathways. Furthermore, gedunin (10 mM) inhibited LPS-induced prostaglandin E 2 production, cyclooxygenase-2 expression, and nuclear factor kB translocation into the nucleus of WT macrophages, demonstrating a wide-range effect of this chemical compound. In addition to the ability to inhibit LPS-induced proinflammatory mediators, gedunin also triggered anti-inflammatory factors interleukin-10, heme oxygenase-1, and Hsp70 in macrophages stimulated or not with LPS. In silico modeling studies revealed that gedunin efficiently docked into the MD-2 LPS binding site, a phenomenon further confirmed by surface plasmon resonance. Our results reveal that, in addition to Hsp90 modulation, gedunin acts as a competitive inhibitor of LPS, blocking the formation of the Toll-like receptor 4/MD-2/LPS complex.

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Section: Introductionmentioning

confidence: 56%

Section: Introductionmentioning

confidence: 99%

Gedunin Binds to Myeloid Differentiation Protein 2 and Impairs Lipopolysaccharide-Induced Toll-Like Receptor 4 Signaling in Macrophages

et al. 2015

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“…In a series of subsequent studies, photoaffinity labeling and proteolysis studies were incorporated into experimentally guided computational predictions to characterize the allosteric C-terminal binding site in Hsp90 at atomic resolution. [198,199] These studies have confirmed that targeted allosteric modulators can antagonize the chaperone function by inducing a conformationally favorable separation of the Hsp90-CTDs and thereby interfere with Hsp90 dimerization. This explained the biological activities of novobiocin and other Hsp90 modulators, providing a new paradigm for the development of allosteric inhibitors to target the C-terminal binding pocket.…”

Section: Hsp90 and Client Proteinsmentioning

confidence: 62%

Computational Studies of Allosteric Regulation in the Hsp90 Molecular Chaperone: From Functional Dynamics and Protein Structure Networks to Allosteric Communications and Targeted Anti‐Cancer Modulators

Verkhivker¹

2014

Israel Journal of Chemistry

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Computational studies of allosteric interactions have witnessed a recent renaissance fueled by growing interest in the modeling of complex molecular assemblies and biological networks. Allosteric interactions of the molecular chaperone Hsp90 with a diverse array of cochaperones and client proteins allow for molecular communication in signal transduction networks. In this review, recent developments in the understanding of allosteric interactions in the context of structural, functional, and computational studies of the Hsp90 chaperone are discussed. A comprehensive analysis of structural and network‐based models of protein allostery is provided. Computational and experimental approaches and advances in the understanding of Hsp90 interactions and regulatory mechanisms are reviewed to provide a systematic and critical view of the current progress and most challenging questions in the field. The current status and future prospects for translational research, bridging the basic science of chaperones with the discovery of anti‐cancer therapies, are also highlighted.

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“…These interact with HSP90 at non-overlapping sites as demonstrated by their ability to capture HSP90 in distinct conformational states [90]. …”

Section: Ligands Targeting the Major Chaperonesmentioning

confidence: 99%

Selective targeting of the stress chaperome as a therapeutic strategy

Taldone

Ochiana

Patel

et al. 2014

Trends in Pharmacological Sciences

105

137

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Normal cellular function is maintained by coordinated proteome machinery that performs a vast array of activities. Helping the proteome in such roles is the chaperome, a network of molecular chaperones and folding enzymes. The stressed cell contains, at any time, a complex mixture of chaperome complexes; a majority performs “housekeeping functions” similarly to non-stressed, normal cells, but a finely-tuned fraction buffers the proteome altered by chronic stress. The stress chaperome is epigenetically distinct from its normal, housekeeping counterpart, providing a basis for its selective targeting by small molecules. Here we discuss development of chaperome inhibitors, and how agents targeting chaperome members in stressed cells are in fact being directed towards chaperome complexes and their effect is therefore determined by their ability to sample and engage such complexes. A new approach is needed to target and implement chaperome modulators in the investigation of diseases, and we propose that the classical thinking in drug discovery needs adjustment when developing chaperome-targeting drugs.

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A systematic protocol for the characterization of Hsp90 modulators

Cited by 78 publications

References 46 publications

Gedunin Binds to Myeloid Differentiation Protein 2 and Impairs Lipopolysaccharide-Induced Toll-Like Receptor 4 Signaling in Macrophages

Gedunin Binds to Myeloid Differentiation Protein 2 and Impairs Lipopolysaccharide-Induced Toll-Like Receptor 4 Signaling in Macrophages

Computational Studies of Allosteric Regulation in the Hsp90 Molecular Chaperone: From Functional Dynamics and Protein Structure Networks to Allosteric Communications and Targeted Anti‐Cancer Modulators

Selective targeting of the stress chaperome as a therapeutic strategy

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