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Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
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Development of a microarray-based assay for efficient testing of new HSP70/DnaK inhibitors
Sona Mohammadi-Ostad-Kalayeh1, Vjaceslavs Hrupins1, Sabine Helmsen1
1Leibniz Universität Hannover, Institute of Biophysics and Center of Biomolecular Drug Research (BMWZ), Schneiderberg 38, D-30167 Hannover, Germany.
Bioorganic & Medicinal Chemistry
|October 19, 2017
Summary
This study introduces a new microarray method for testing ATP binding, crucial for understanding chaperone protein function. The method identifies novel HSP70 inhibitors from natural products and analyzes protein interactions.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- Heat shock proteins (HSPs) are vital molecular chaperones involved in protein homeostasis.
- ATP binding is central to HSP function, making it a key target for therapeutic intervention.
- Developing efficient methods to study ATP binding and inhibition is crucial for drug discovery.
Purpose of the Study:
- To develop a facile, miniaturized microarray method for assessing ATP binding to HSP70.
- To investigate the impact of ATP labeling position on binding affinity.
- To screen for novel HSP70 inhibitors and analyze protein-protein interactions.
Main Methods:
- Utilized a miniaturized microarray platform for ATP binding assays.
- Employed human HSP70 and Mycobacterium tuberculosis DnaK as biological targets.
- Incorporated molecular modeling to understand label positioning effects.
- Screened a natural product library and analyzed protein-protein interactions.
Main Results:
- Demonstrated that ATP's fluorescence label position significantly affects HSP70 binding, with optimal binding occurring when labeled on the adenine ring.
- Identified terphenyls rickenyl A and D from Hypoxylon rickii as novel HSP70 inhibitors with IC50 values in the micromolar range.
- Quantified the affinity of HSP90 for HSP70, revealing a significant protein-protein interaction.
Conclusions:
- The developed microarray assay is a robust and versatile tool for ATP binding studies, inhibitor screening, and protein interaction analysis.
- The findings highlight the importance of ATP labeling strategy in biochemical assays.
- Identified new natural product inhibitors of HSP70, offering potential leads for therapeutic development.
