Microarray-based screening of heat shock protein inhibitors

verfasst von

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.

Organisationseinheit(en)

Institut für Technische Chemie
Zentrum für Biomolekulare Wirkstoffe (BMWZ)
Institut für Organische Chemie
Abteilung Zellphysiologie und Biophysik

Externe Organisation(en)

University of California at San Francisco

Typ

Artikel

Journal

Journal of biotechnology

Band

180

Seiten

1-9

Anzahl der Seiten

9

ISSN

0168-1656

Publikationsdatum

22.03.2014

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Biotechnologie, Bioengineering, Angewandte Mikrobiologie und Biotechnologie

Ziele für nachhaltige Entwicklung

SDG 3 – Gute Gesundheit und Wohlergehen

Elektronische Version(en)

https://doi.org/10.1016/j.jbiotec.2014.03.006 (Zugang: Geschlossen)