The Novolactone Natural Product Disrupts the Allosteric Regulation of Hsp70

Hassan, Amr; Kirby, C.A.; Zhou, Wenlai; Schuhmann, Tim; Kityk, Roman; Kipp, D. Randal; Baird, Jason R.; Chen, Jinyun; Chen, Yaoyu; Chung, Franklin; Hoepfner, Dominic; Movva, N. Rao; Pagliarini, Raymond; Petersen, Frank; Quinn, Christopher; Quinn, Douglas; Riedl, Ralph; Schmitt, E.; Schitter, Anne; Stams, Travis; Studer, Christian; Fortin, Pascal D.; Mayer, Mathias; Sadlish, Heather

doi:10.1016/j.chembiol.2014.11.007

Cited by 50 publications

(87 citation statements)

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“…Even minor perturbations in these hotspots propagate through the entire βSBD, suggesting how substrate-binding and nucleotide-induced domain docking can regulate Hsp70 functions. Strikingly, the recent discovery of new allosteric inhibitors of Hsp70s (26,37) demonstrates that, indeed, the region around the β5/β7/β8 hydrophobic core is a promising druggable site, fully in line with our results. Moreover, the exquisite sensitivity to subtle changes in these βSBD allosteric hotspots suggests how evolutionary tuning of Hsp70s and modulation by posttranslational modifications and cochaperone interactions can alter Hsp70 functional behavior.…”

Section: Discussionsupporting

confidence: 80%

Substrate-binding domain conformational dynamics mediate Hsp70 allostery

Zhuravleva

Gierasch

2015

Proc. Natl. Acad. Sci. U.S.A.

105

164

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Binding of ATP to the N-terminal nucleotide-binding domain (NBD) of heat shock protein 70 (Hsp70) molecular chaperones reduces the affinity of their C-terminal substrate-binding domain (SBD) for unfolded protein substrates. ATP binding to the NBD leads to docking between NBD and βSBD and releasing of the α-helical lid that covers the substrate-binding cleft in the SBD. However, these structural changes alone do not fully account for the allosteric mechanism of modulation of substrate affinity and binding kinetics. Through a multipronged study of the Escherichia coli Hsp70 DnaK, we found that changes in conformational dynamics within the βSBD play a central role in interdomain allosteric communication in the Hsp70 DnaK. ATP-mediated NBD conformational changes favor formation of NBD contacts with lynchpin sites on the βSBD and force disengagement of SBD strand β8 from strand β7, which leads to repacking of a βSBD hydrophobic cluster and disruption of the hydrophobic arch over the substrate-binding cleft. In turn, these structural rearrangements drastically enhance conformational dynamics throughout the entire βSBD and particularly around the substrate-binding site. This negative, entropically driven allostery between two functional sites of the βSBD-the NBD binding interface and the substrate-binding site-confers upon the SBD the plasticity needed to bind to a wide range of chaperone clients without compromising precise control of thermodynamics and kinetics of chaperone-client interactions. molecular chaperone | protein quality control | NMR chemical shift perturbations | conformational selection | entropically driven allostery H eat shock protein 70 (Hsp70) molecular chaperones are central players in protein quality control systems for organisms from bacteria to humans (1, 2). All Hsp70s consist of two highly conserved domains: an N-terminal nucleotide-binding domain (NBD), which regulates the affinity of substrate binding, and a C-terminal substrate-binding domain (SBD), which binds to exposed hydrophobic stretches of client proteins and is made up of a β-sandwich domain (the βSBD), and an α-helical lid (the αLid) (Fig. 1A). Hsp70 functions rely on interdomain allostery: ATP hydrolysis in the NBD controls thermodynamics and kinetics of substrate binding and release; in turn, substrate binding to the SBD stimulates ATPase activity in the NBD (3-5).Mechanistic understanding of Hsp70 function has been greatly enhanced by recent breakthrough achievements in structural characterization of individual functional steps of the allosteric cycle for the Escherichia coli Hsp70 DnaK. New crystal structures and NMR analysis of DnaK have provided descriptions of three major functional states: ADP-bound (6, 7), 9), and ATP/substrate-bound (10), which have distinct arrangements of the four structural units NBD, βSBD, αLid, and interdomain linker (Fig. 1A). In the ADP-bound (domain-undocked, linker-unbound) state, the two DnaK domains, the NBD and SBD, behave independently, with the interdomain linker exposed to the solvent and ...

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Section: Discussionsupporting

confidence: 80%

Substrate-binding domain conformational dynamics mediate Hsp70 allostery

Zhuravleva

Gierasch

2015

Proc. Natl. Acad. Sci. U.S.A.

105

164

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“…1b) [23]. Obviously, best applicability of this assay is in the area of antifungal research but due to significant biological conservation the HIP methodology supported target identification of several targets also in other species including Plasmodium and human as presented in this study by cladosporin [24], novolactone [19] and rocaglamide [49]. Additional examples are shown in the study by Hoepfner et al [23].…”

Section: Methods For Target Identificationmentioning

confidence: 66%

Natural products as probes in pharmaceutical research

Schmitt

Hoepfner

Krastel

2016

Journal of Industrial Microbiology and Biotechnology

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From the start of the pharmaceutical research natural products played a key role in drug discovery and development. Over time many discoveries of fundamental new biology were triggered by the unique biological activity of natural products. Unprecedented chemical structures, novel chemotypes, often pave the way to investigate new biology and to explore new pathways and targets. This review summarizes the recent results in the area with a focus on research done in the laboratories of Novartis Institutes for BioMedical Research. We aim to put the technological advances in target identification techniques in the context to the current revival of phenotypic screening and the increasingly complex biological questions related to drug discovery.

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“…A group at Novartis recently described another inhibitor that binds in the SBD [124]. The authors started with a genomic approach to identify natural products with activity on Hsp70.…”

Section: Novolactonementioning

confidence: 99%

Allosteric Inhibitors of Hsp70: Drugging the Second Chaperone of Tumorigenesis

Srinivasan

Shao

et al. 2015

Topics in Medicinal Chemistry

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Cancer cells survive in the presence of stresses that would normally cause cell death. To accomplish this feat, they express elevated levels of the molecular chaperones: heat shock protein 70 (Hsp70) and heat shock protein 90 (Hsp90). Knockdown of these chaperones is selectively toxic to cancer cells, suggesting that they might be promising nodes for anticancer therapy. However, while inhibitors of Hsp90 are well known, progress in the development of Hsp70 inhibitors has proven more difficult. Hsp70 binds tightly to ATP through a highly conserved domain of the actin/hexokinase superfamily, making it challenging to identify selective, competitive inhibitors. Despite this obstacle, progress has been made and first-generation molecules are being deployed. To supplement these efforts, compounds that target important allosteric sites on the chaperone have also been discovered. In some of these cases, the molecules have been shown to control key protein-protein interactions between Hsp70 and its co-chaperones. In other cases, allosteric sites have been used to gain unexpected selectivity for members of the Hsp70 family. Here, we review recent progress in the development of Hsp70 inhibitors.

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The Novolactone Natural Product Disrupts the Allosteric Regulation of Hsp70

Cited by 50 publications

References 50 publications

Substrate-binding domain conformational dynamics mediate Hsp70 allostery

Substrate-binding domain conformational dynamics mediate Hsp70 allostery

Natural products as probes in pharmaceutical research

Allosteric Inhibitors of Hsp70: Drugging the Second Chaperone of Tumorigenesis

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