Microarray-based screening of heat shock protein inhibitors

authored by: Emilia Schax, Johanna Gabriela Walter, Helene Märzhäuser, Frank Stahl, Thomas Scheper, David A. Agard, Simone Eichner, Andreas Kirschning, Carsten Zeilinger
Abstract: Based on the importance of heat shock proteins (HSPs) in diseases such as cancer, Alzheimer's disease or malaria, inhibitors of these chaperons are needed. Today's state-of-the-art techniques to identify HSP inhibitors are performed in microplate format, requiring large amounts of proteins and potential inhibitors. In contrast, we have developed a miniaturized protein microarray-based assay to identify novel inhibitors, allowing analysis with 300pmol of protein. The assay is based on competitive binding of fluorescence-labeled ATP and potential inhibitors to the ATP-binding site of HSP. Therefore, the developed microarray enables the parallel analysis of different ATP-binding proteins on a single microarray. We have demonstrated the possibility of multiplexing by immobilizing full-length human HSP90α and HtpG of Helicobacter pylori on microarrays. Fluorescence-labeled ATP was competed by novel geldanamycin/reblastatin derivatives with IC₅₀ values in the range of 0.5nM to 4μM and Z^*-factors between 0.60 and 0.96. Our results demonstrate the potential of a target-oriented multiplexed protein microarray to identify novel inhibitors for different members of the HSP90 family.
Organisation(s): Institute of Technical Chemistry
Centre of Biomolecular Drug Research (BMWZ)
Institute of Organic Chemistry
Department of Cell Physiology and Biophysics
External Organisation(s): University of California at San Francisco
Type: Article
Journal: Journal of biotechnology
Volume: 180
Pages: 1-9
No. of pages: 9
ISSN: 0168-1656
Publication date: 22.03.2014
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Biotechnology, Bioengineering, Applied Microbiology and Biotechnology
Sustainable Development Goals: SDG 3 - Good Health and Well-being
Electronic version(s): https://doi.org/10.1016/j.jbiotec.2014.03.006 (Access: Closed)