Medium-Throughput Detection of Hsp90/Cdc37 Protein–Protein Interaction Inhibitors Using a Split Renilla Luciferase-Based Assay

A1 Originalartikel i en vetenskaplig tidskrift (referentgranskad)

Interna författare/redaktörer

Publikationens författare: Farid Ahmad Siddiqui, Hanna Parkkola, Ganesh babu Manoharan, Daniel Abankwa
Förläggare: Sage
Publiceringsår: 2020
Tidskrift: Slas Discovery
Volym: 25
Nummer: 2
Artikelns första sida, sidnummer: 195
Artikelns sista sida, sidnummer: 206


The protein-folding chaperone Hsp90 enables the maturation and stability of various oncogenic signaling proteins and is
thus pursued as a cancer drug target. Folding in particular of protein kinases is assisted by the co-chaperone Cdc37. Several
inhibitors against the Hsp90 ATP-binding site have been developed. However, they displayed significant toxicity in clinical
trials. By contrast, the natural product conglobatin A has an exceptionally low toxicity in mice. It targets the protein–protein
interface (PPI) of Hsp90 and Cdc37, suggesting that interface inhibitors have an interesting drug development potential.

In order to identify inhibitors of the Hsp90/Cdc37 PPI, we have established a mammalian cell lysate-based, medium-
throughput amenable split Renilla luciferase assay. This assay employs N-terminal and C-terminal fragments of Renilla
luciferase fused to full-length human Hsp90 and Cdc37, respectively. We expect that our assay will allow for the
identification of novel Hsp90/Cdc37 interaction inhibitors. Such tool compounds will help to evaluate whether the toxicity
profile of Hsp90/Cdc37 PPI inhibitors is in general more favorable than that of ATP-competitive Hsp90 inhibitors. Further
development of such tool compounds may lead to new classes of Hsp90 inhibitors with applications in cancer and other

Senast uppdaterad 2020-29-05 vid 06:57