Hsp90-Tau Complex Reveals Molecular Basis for Specificity in Chaperone Action

“Spectra were processed with NMRPipe (Delaglio et al., 1995) and analyzed using CcpNMR Analysis (Vranken et al., 2005).”

“The Hsp90-Tau Interaction and Therapeutic Treatment of Alzheimer Disease Aggregation of the Tau microtubule binding region causes Alzheimer disease (Haass and Selkoe, 2007; Kosik et al., 1986).”

“in Chaperone Action G. Elif Karagöz,1,2 Afonso M.S. Duarte,1,3 Elias Akoury,4 Hans Ippel,5,6 Jacek Biernat,7 Tania Morán Luengo,1 Martina Radli,1 Tatiana Didenko,1,15 Bryce A. Nordhues,8 Dmitry B. Veprintsev,9 Chad A. Dickey,8 EckhardMandelkow,7,10 Markus Zweckstetter,4,11,12 Rolf Boelens,5 Tobias Madl,5,13,14,* and…”

“Spectra were processed with NMRPipe (Delaglio et al., 1995) and analyzed using CcpNMR Analysis (Vranken et al., 2005).”

“Recent work suggests that Tau amyloid formation may actually be a protectivemechanism that allows potentially toxic Tau intermediates to self-chaperone and sequester themselves into less toxic entities within long-lived neurons (Eisenberg and Jucker, 2012; Oddo et al., 2004; Santacruz et al., 2005).”

“It also accumulates abnormally in brain tissue of Alzheimer disease patients and is closely linkedwith neuronal toxicity in this most commonneurodegenerative disease (Ittner et al., 2010; Roberson et al., 2007).”

“It also accumulates abnormally in brain tissue of Alzheimer disease patients and is closely linkedwith neuronal toxicity in this most commonneurodegenerative disease (Ittner et al., 2010; Roberson et al., 2007).”

“The substrate pool of Hsp90 does not share a common sequence or structural motif (Taipale et al., 2010, 2012).”

“Crucial for the chaperone network in the cytoplasm are the ATP-driven Hsp70 and Hsp90 families (Bukau et al., 2006; Mayer, 2010).”

“The Hsp90-Tau Interaction and Therapeutic Treatment of Alzheimer Disease Aggregation of the Tau microtubule binding region causes Alzheimer disease (Haass and Selkoe, 2007; Kosik et al., 1986).”

“…provides hydrophobic interaction and negative charges similar to the other chaperone systems, it also provides a dramatically more extended substrate-binding site than any of the other chaperones (Figure 5) (Buckle et al., 1997; Rüdiger et al., 1997a; Weibe- zahn et al., 2005; Xu et al., 1997).”

“Hsp90 interacts with a specific, but structurally diverse, subset of client Cell 156, 963–974, February 27, 2014 ª2014 Elsevier Inc. 963 proteins including kinases and steroid receptors (Echeverria and Picard, 2010; Jakob et al., 1995; Pearl and Prodromou, 2006; Street et al., 2011).”

“The Hsp90-binding region contains the entire microtubulebinding repeats domain, including the 275VQIINK280 and 306VQIVYK311 hexapeptide motifs, which are crucial for pathological Tau aggregation in Alzheimer disease (Figures 1E and 1F) (von Bergen et al., 2000).”

“We combined the structural information that could be extracted from our NMR restraints and scattering curves with previously published homology models based on published crystal structures of Hsp90 (Ali et al., 2006; Karagöz et al., 2011).”

“Tau normally stabilizes microtubules that serve as tracks for axonal transport in neurons (Mandelkow and Mandelkow, 2012).”

“SAXSbased structural models were generated using the program CORAL (Petoukhov and Svergun, 2005), known crystal structures, a homology model for Hsp90 (Karagöz et al.”

“The substrate pool of Hsp90 does not share a common sequence or structural motif (Taipale et al., 2010, 2012).”

“Full-length N-labeled human Tau protein (hTau40) was expressed and purified as described previously (Mukrasch et al., 2009).”

“Spectra processing, analysis, and representations were done as described (Karagöz et al., 2011; Tugarinov et al., 2003).”

“Perhaps by trying to chaperone the microtubule binding domains of Tau, Hsp90 and other chaperones are extending the half-life of the disordered component in the cell, increasing its potential for toxicity (Dickey et al., 2007; Jinwal et al., 2013).”

“Hsp90 contributes to folding and maturation in concert with other chaperone systems, in particular Hsp70 (Wegele et al., 2004).”

“The data were subjected to nonlinear least-square fitting using the Solver module in Excel (Kemmer and Keller, 2010), according to the equation”

“…caused by the combined effects of (1) the increase in molecular weight upon complex formation, which increases transverse relaxation rates and thereby line width, and (2) the chemical exchange at the contact surface (Dekker et al., 1993; Sette et al., 1997; Wüthrich, 2003; Zuiderweg, 2002).”

“Therefore, we mapped the binding sites of several key cochaperones, including Aha1, p23, Cdc37, Sgt-1, and CHORD, which have previously been found in cocrystals with Hsp90 (Figure 5E) (Li et al., 2012; Pearl et al., 2008).”

“We noted, however, that the Tau-binding sites in both Hsp90N and Hsp90-M have been implicated in early crystal structures of both domains as potential substrate-binding sites (Figure 4) (Meyer et al., 2003; Prodromou et al., 1997).”

“Mutant Tau is particularly susceptible to inhibition of Hsp90, making it a promising lead for therapeutic strategies in Alzheimer disease and other Tau-related disorders (Dou et al., 2003; Luo et al., 2007).”

“Hsp90 functions as a dynamic dimer that is characterized by large interdomain movements, which is also reflected by the crystal structures of Hsp90 homologs captured in different conformations (Ali et al., 2006; Dollins et al., 2007; Shiau et al., 2006).”

“Therefore, we mapped the binding sites of several key cochaperones, including Aha1, p23, Cdc37, Sgt-1, and CHORD, which have previously been found in cocrystals with Hsp90 (Figure 5E) (Li et al., 2012; Pearl et al., 2008).”

“During the folding process, these short hydrophobic stretches will most likely form the nucleus around which the protein condenses (Daggett and Fersht, 2003).”

“…provides hydrophobic interaction and negative charges similar to the other chaperone systems, it also provides a dramatically more extended substrate-binding site than any of the other chaperones (Figure 5) (Buckle et al., 1997; Rüdiger et al., 1997a; Weibe- zahn et al., 2005; Xu et al., 1997).”

“Crucial for the chaperone network in the cytoplasm are the ATP-driven Hsp70 and Hsp90 families (Bukau et al., 2006; Mayer, 2010).”

“Hsp90 interacts with a specific, but structurally diverse, subset of client Cell 156, 963–974, February 27, 2014 ª2014 Elsevier Inc. 963 proteins including kinases and steroid receptors (Echeverria and Picard, 2010; Jakob et al., 1995; Pearl and Prodromou, 2006; Street et al., 2011).”

“As recently described, Hsp90 and the cochaperone FKBP51 can prevent Tau from forming dense amyloids and rather promote the formation of more toxic Tau intermediates (Blair et al., 2013).”

“While there is no evidence to support the first possibility, conformational asymmetry is characteristic for several previously described Hsp90 complexes (i.e., Hsp90-p23, Hsp90-Cdk4-Cdc37, and Hsp90Aha1) (Karagöz et al., 2011; Retzlaff et al., 2010; Vaughan et al., 2006).”

“Hsp90 functions as a dynamic dimer that is characterized by large interdomain movements, which is also reflected by the crystal structures of Hsp90 homologs captured in different conformations (Ali et al., 2006; Dollins et al., 2007; Shiau et al., 2006).”

“This domain differs in sequence composition from the typical IDP by a higher content in hydrophobics, a typical trigger for chaperone interaction (Figure 1E) (Dunker and Gough, 2011; Hartl et al., 2011; Uversky et al., 2009).”

“…caused by the combined effects of (1) the increase in molecular weight upon complex formation, which increases transverse relaxation rates and thereby line width, and (2) the chemical exchange at the contact surface (Dekker et al., 1993; Sette et al., 1997; Wüthrich, 2003; Zuiderweg, 2002).”

“While there is no evidence to support the first possibility, conformational asymmetry is characteristic for several previously described Hsp90 complexes (i.e., Hsp90-p23, Hsp90-Cdk4-Cdc37, and Hsp90Aha1) (Karagöz et al., 2011; Retzlaff et al., 2010; Vaughan et al., 2006).”

“The border on one side is the long helix mH2 and on the other side it is the amphipathic loop around Leu342/Phe343, which has been earlier implicated in client interaction (Prodromou et al., 1997).”

“Hsp90 interacts with a specific, but structurally diverse, subset of client Cell 156, 963–974, February 27, 2014 ª2014 Elsevier Inc. 963 proteins including kinases and steroid receptors (Echeverria and Picard, 2010; Jakob et al., 1995; Pearl and Prodromou, 2006; Street et al., 2011).”

“were produced and purified as described (Barghorn et al., 2004; Karagöz et al., 2011).”

“…higher radius of gyration and a larger maximal extension than expected for globular proteins of the same size, consistent with its disordered nature and with previous studies (Rg = 6.8 ± 1.8 nm, Dmax = 24.9 nm; Figures 3B and 3C; Table 1) (Mylonas et al., 2008; Shkumatov et al., 2011).”

“…provides hydrophobic interaction and negative charges similar to the other chaperone systems, it also provides a dramatically more extended substrate-binding site than any of the other chaperones (Figure 5) (Buckle et al., 1997; Rüdiger et al., 1997a; Weibe- zahn et al., 2005; Xu et al., 1997).”

“…of South Florida Health Byrd Alzheimer’s Institute, University of South Florida, Tampa, FL 33613, USA 9Laboratory of Biomolecular Research, Paul Scherrer Institut, 5232 Villigen PSI, Switzerland and Department of Biology, ETH Zürich, 8093 Zürich, Switzerland 10CAESAR Research Center,…”

“We relate this to the fact that ATP binding alone does not induce stable conformational changes in human Hsp90, in contrast to the yeast and E. coli homologs (Figure S2) (Krukenberg et al., 2008, 2009).”

“…caused by the combined effects of (1) the increase in molecular weight upon complex formation, which increases transverse relaxation rates and thereby line width, and (2) the chemical exchange at the contact surface (Dekker et al., 1993; Sette et al., 1997; Wüthrich, 2003; Zuiderweg, 2002).”

“…caused by the combined effects of (1) the increase in molecular weight upon complex formation, which increases transverse relaxation rates and thereby line width, and (2) the chemical exchange at the contact surface (Dekker et al., 1993; Sette et al., 1997; Wüthrich, 2003; Zuiderweg, 2002).”

“This feature is not reserved for Tau alone, but for other disease-associated proteins as well, such as a-synuclein (Daturpalli et al., 2013).”

“Studies that used isolated domains and/or worked at subphysiological salt conditions also implied other surface elements involved in substrate binding (Didenko et al., 2012; Hagn et al., 2011; Park et al., 2011).”

“This allows recognition of scattered hydrophobic residues in late folding intermediates that remain after early burial of the Hsp70 sites.”

“specifically bind to two short segments within this region (VQIIN and VQIVY; Figures 1C and 1F) (Jinwal et al., 2013; Sarkar et al., 2008).”

“For the E. coli Hsp90, some mutations in Hsp90-M and Hsp90-C were recently implicated in substrate-binding, and in fact, the mutations in Hsp90-M are close to the Tau site (Genest et al., 2013).”

“…caused by the combined effects of (1) the increase in molecular weight upon complex formation, which increases transverse relaxation rates and thereby line width, and (2) the chemical exchange at the contact surface (Dekker et al., 1993; Sette et al., 1997; Wüthrich, 2003; Zuiderweg, 2002).”

“The repeat region is indicated in cyan. specifically bind to two short segments within this region (275VQIIN279 and 306VQIVY310; Figures 1C and 1F) (Jinwal et al., 2013; Sarkar et al., 2008).”

“Studies that used isolated domains and/or worked at subphysiological salt conditions also implied other surface elements involved in substrate binding (Didenko et al., 2012; Hagn et al., 2011; Park et al., 2011).”

“Interestingly, Hsp90 also interacts with some intrinsically disordered proteins, including the microtubule-associated protein Tau (Dickey et al., 2007; Dou et al., 2003; Miyata et al., 2011; Thompson et al., 2012).”

“Studies that used isolated domains and/or worked at subphysiological salt conditions also implied other surface elements involved in substrate binding (Didenko et al., 2012; Hagn et al., 2011; Park et al., 2011).”

“…higher radius of gyration and a larger maximal extension than expected for globular proteins of the same size, consistent with its disordered nature and with previous studies (Rg = 6.8 ± 1.8 nm, Dmax = 24.9 nm; Figures 3B and 3C; Table 1) (Mylonas et al., 2008; Shkumatov et al., 2011).”

“…and Department of Biology, ETH Zürich, 8093 Zürich, Switzerland 10CAESAR Research Center, Ludwig-Erhard-Allee 2, 53175 Bonn, Germany 11German Center for Neurodegenerative Diseases (DZNE), 37077 Göttingen, Germany 12Center for Nanoscale Microscopy and Molecular Physiology of the Brain…”

“We relate this to the fact that ATP binding alone does not induce stable conformational changes in human Hsp90, in contrast to the yeast and E. coli homologs (Figure S2) (Krukenberg et al., 2008, 2009).”

“Isoleucines are not involved in hydrogen bonds and therefore less sensitive to long-range structural changes when compared to backbone amide signals, as observed in previous isoleucine-based NMR analyses of Hsp90 complexes (Didenko et al., 2011, 2012; Karagöz et al., 2011).”

“This domain differs in sequence composition from the typical IDP by a higher content in hydrophobics, a typical trigger for chaperone interaction (Figure 1E) (Dunker and Gough, 2011; Hartl et al., 2011; Uversky et al., 2009).”

“Recent evidence suggests that Hsp90 has an extended substrate binding interface that crosses domain boundaries and exhibits specificity for proteins with exposed hydrophobic residues, independent of whether they are disordered, partly folded, or near native (113, 117, 118).”