Document Detail


Identification of small molecules that modify the protein folding activity of heat shock protein 70.
MedLine Citation:
PMID:  18191466     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Molecular chaperones, such as heat shock protein 70 (Hsp70) and its bacterial ortholog DnaK, play numerous important roles in protein folding. In vitro, this activity can be observed by incubating purified chaperones with denatured substrates and measuring the recovery of properly folded protein. In an effort to rapidly identify small molecules that modify this folding activity, we modified an existing method for use in 96-well plates. In this assay, denatured firefly luciferase was treated with a mixture of DnaK and prospective chemical modulators. The luminescence of refolded luciferase was used to follow the reaction progress, and counterscreens excluded compounds that target luciferase; thus, hits from these screens modify protein folding via their effects on the function of the chaperone machine. Using this platform, we screened a pilot chemical library and found five new inhibitors of DnaK and one compound that promoted folding. These chemical probes may be useful in studies aimed at understanding the many varied roles of chaperones in cellular protein folding. Moreover, this assay provides the opportunity to rapidly screen for additional compounds that might regulate the folding activity of Hsp70.
Authors:
Susanne Wisén; Jason E Gestwicki
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2007-12-14
Journal Detail:
Title:  Analytical biochemistry     Volume:  374     ISSN:  1096-0309     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-02-12     Completed Date:  2008-06-24     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  371-7     Citation Subset:  IM    
Affiliation:
Departments of Pathology and Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, USA.
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphate / metabolism
Chemistry Techniques, Analytical / instrumentation
Drug Evaluation, Preclinical*
HSP40 Heat-Shock Proteins / metabolism
HSP70 Heat-Shock Proteins / metabolism*
Luciferases, Firefly / metabolism
Protein Folding*
Protein Renaturation / drug effects
Sensitivity and Specificity
Small Molecule Libraries / pharmacology*
Grant Support
ID/Acronym/Agency:
P50/GM067082/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/HSP40 Heat-Shock Proteins; 0/HSP70 Heat-Shock Proteins; 0/Small Molecule Libraries; 56-65-5/Adenosine Triphosphate; EC 1.13.12.7/Luciferases, Firefly

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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