title: The novolactone natural product disrupts the allosteric regulation of Hsp70.
abstract: The highly conserved 70 kDa heat shock proteins (Hsp70) play an integral role in proteostasis such that dysregulation has been implicated in numerous diseases. Elucidating the precise role of Hsp70 family members in the cellular context, however, has been hampered by the redundancy and intricate regulation of the chaperone network, and relatively few selective and potent tools. We have characterized a natural product, novolactone, that targets cytosolic and ER-localized isoforms of Hsp70 through a highly conserved covalent interaction at the interface between the substrate-binding and ATPase domains. Biochemical and structural analyses indicate that novolactone disrupts interdomain communication by allosterically inducing a conformational change in the Hsp70 protein to block ATP-induced substrate release and inhibit refolding activities. Thus, novolactone is a valuable tool for exploring the requirements of Hsp70 chaperones in diverse cellular contexts.
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: A Quamrul
authors.initials: AQ
authors.lastname: Hassan
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Christina A
authors.initials: CA
authors.lastname: Kirby
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Wenlai
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Tim
authors.initials: T
authors.lastname: Schuhmann
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
authors.forename: Roman
authors.initials: R
authors.lastname: Kityk
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: D Randal
authors.initials: DR
authors.lastname: Kipp
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Baird
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Jinyun
authors.initials: J
authors.lastname: Chen
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Yaoyu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Franklin
authors.initials: F
authors.lastname: Chung
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Dominic
authors.initials: D
authors.lastname: Hoepfner
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: N Rao
authors.initials: NR
authors.lastname: Movva
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Raymond
authors.initials: R
authors.lastname: Pagliarini
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Frank
authors.initials: F
authors.lastname: Petersen
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Quinn
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Douglas
authors.initials: D
authors.lastname: Quinn
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Ralph
authors.initials: R
authors.lastname: Riedl
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Esther K
authors.initials: EK
authors.lastname: Schmitt
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Anne
authors.initials: A
authors.lastname: Schitter
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Travis
authors.initials: T
authors.lastname: Stams
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Studer
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Pascal D
authors.initials: PD
authors.lastname: Fortin
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Mayer
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland. Electronic address: heather.sadlish@novartis.com.
authors.forename: Heather
authors.initials: H
authors.lastname: Sadlish
mesh_terms: D045784:Abietanes / Q000737:chemistry / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000494:Allosteric Regulation; D001665:Binding Sites; D001688:Biological Products / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D018360:Crystallography, X-Ray; D004721:Endoplasmic Reticulum / Q000378:metabolism; D016681:Genome, Fungal; D050956:HSP40 Heat-Shock Proteins / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D045784:Abietanes; D001688:Biological Products; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D020033:Protein Isoforms; C000601548:novolactone; D000251:Adenosine Triphosphatases