ChemBioChem · 2010 · 14 citations · 50 references
Combinatorial ChemistryE. ColiProtein AssemblyMolecular BiologySecondary AmideHsp70 Chaperone DnakProtein FoldingRational DesignProteomicsProtein FunctionBiochemistryBiomolecular InteractionStructural BiologyBiomolecular EngineeringNatural SciencesPeptide LibraryPeptoidProtein EngineeringMolecular ComplexMedicine
The hsp70 chaperone DnaK from E. coli plays a major role in cellular stress response and is involved in assisted protein folding in vivo. By screening a combinatorial peptide library, we identified several DnaK-specific peptide ligands with nanomolar affinities, which are able to inhibit the secondary amide peptide bond cis/trans isomerase (APIase) activity of DnaK, as well as DnaK/DnaJ/GrpE-assisted refolding of firefly luciferase. Our designed DnaK inhibitors have the capability to penetrate E. coli cells and feature a high protease resistance. Once inside the cell, they physically target DnaK. NMR-based (1)H/(15)N-HSQC experiments furthermore confirmed that the designed peptidic ligands all bind in an identical manner to the conventional peptide-binding site of DnaK. The subsequent blocking of DnaK function apparently results in the observed antibacterial effects on E. coli cells, with minimum inhibitory concentrations in the range of 100 microM.
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Structural Analysis of Substrate Binding by the Molecular Chaperone DnaK
Xiaotian Zhu, Xun Zhao, William F. Burkholder et al. · Science · 1996 · 1.3K citations