eprintid: 18794 rev_number: 21 userid: 1419 dir: disk0/00/01/87/94 datestamp: 2013-05-29 16:32:56 lastmod: 2019-02-13 12:55:10 status_changed: 2013-05-29 16:32:56 type: article metadata_visibility: show item_issues_count: 0 eprint_status: archive creators_name: Wisén, S creators_name: Bertelsen, EB creators_name: Thompson, AD creators_name: Patury, S creators_name: Ung, P creators_name: Chang, L creators_name: Evans, CG creators_name: Walter, GM creators_name: Wipf, P creators_name: Carlson, HA creators_name: Brodsky, JL creators_name: Zuiderweg, ERP creators_name: Gestwicki, JE creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: pwipf@pitt.edu creators_email: creators_email: jbrodsky@pitt.edu creators_email: creators_email: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: PWIPF creators_id: creators_id: JBRODSKY creators_id: creators_id: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: creators_orcid: 0000-0002-6984-8486 creators_orcid: creators_orcid: title: Binding of a small molecule at a protein-protein interface regulates the chaperone activity of Hsp70-Hsp40 ispublished: pub divisions: sch_as_chemistry full_text_status: public abstract: Heat shock protein 70 (Hsp70) is a highly conserved molecular chaperone that plays multiple roles in protein homeostasis. In these various tasks, the activity of Hsp70 is shaped by interactions with co-chaperones, such as Hsp40. The Hsp40 family of co-chaperones binds to Hsp70 through a conserved J-domain, and these factors stimulate ATPase and protein-folding activity. Using chemical screens, we identified a compound, 115-7c, which acts as an artificial co-chaperone for Hsp70. Specifically, the activities of 115-7c mirrored those of a Hsp40; the compound stimulated the ATPase and protein-folding activities of a prokaryotic Hsp70 (DnaK) and partially compensated for a Hsp40 loss-of-function mutation in yeast. Consistent with these observations, NMR and mutagenesis studies indicate that the binding site for 115-7c is adjacent to a region on DnaK that is required for J-domain-mediated stimulation. Interestingly, we found that 115-7c and the Hsp40 do not compete for binding but act in concert. Using this information, we introduced additional steric bulk to 115-7c and converted it into an inhibitor. Thus, these chemical probes either promote or inhibit chaperone functions by regulating Hsp70-Hsp40 complex assembly at a native protein-protein interface. This unexpected mechanism may provide new avenues for exploring how chaperones and co-chaperones cooperate to shape protein homeostasis. © 2010 American Chemical Society. date: 2010-06-18 date_type: published publication: ACS Chemical Biology volume: 5 number: 6 pagerange: 611 - 622 refereed: TRUE issn: 1554-8929 id_number: 10.1021/cb1000422 other_id: NLM NIHMS240226 other_id: NLM PMC2950966 pmcid: PMC2950966 pmid: 20481474 mesh_headings: Escherichia coli--chemistry mesh_headings: Escherichia coli--genetics mesh_headings: Escherichia coli--metabolism mesh_headings: Escherichia coli Proteins--chemistry mesh_headings: Escherichia coli Proteins--genetics mesh_headings: Escherichia coli Proteins--metabolism mesh_headings: Gene Expression Regulation, Fungal--drug effects mesh_headings: HSP40 Heat-Shock Proteins--metabolism mesh_headings: HSP70 Heat-Shock Proteins--chemistry mesh_headings: HSP70 Heat-Shock Proteins--genetics mesh_headings: HSP70 Heat-Shock Proteins--metabolism mesh_headings: Models, Molecular mesh_headings: Mutagenesis, Site-Directed mesh_headings: Protein Binding mesh_headings: Protein Structure, Tertiary mesh_headings: Saccharomyces cerevisiae--genetics mesh_headings: Saccharomyces cerevisiae--metabolism mesh_headings: Saccharomyces cerevisiae Proteins--genetics mesh_headings: Saccharomyces cerevisiae Proteins--metabolism mesh_headings: Small Molecule Libraries--chemistry mesh_headings: Small Molecule Libraries--pharmacology chemical_names: Escherichia coli Proteins chemical_names: HSP40 Heat-Shock Proteins chemical_names: HSP70 Heat-Shock Proteins chemical_names: Saccharomyces cerevisiae Proteins chemical_names: Small Molecule Libraries chemical_names: dnaK protein, E coli citation: Wisén, S and Bertelsen, EB and Thompson, AD and Patury, S and Ung, P and Chang, L and Evans, CG and Walter, GM and Wipf, P and Carlson, HA and Brodsky, JL and Zuiderweg, ERP and Gestwicki, JE (2010) Binding of a small molecule at a protein-protein interface regulates the chaperone activity of Hsp70-Hsp40. ACS Chemical Biology, 5 (6). 611 - 622. ISSN 1554-8929 document_url: http://d-scholarship-dev.library.pitt.edu/18794/1/licence.txt