Abstract
Kinase enzymes are an important class of drug targets, particularly in cancer. Cell-based kinase assays are needed to understand how potential kinase inhibitors act on their targets in a physiologically relevant context. Current cell-based kinase assays rely on antibodybased detection of endogenous substrates, inaccurate disease models, or indirect measurements of drug action. Here we expand on previous work from our lab to introduce a 96-well plate compatible approach for measuring cell-based kinase activity in disease-relevant human chronic myeloid leukemia cell lines using an exogenously added, multi-functional peptide substrate. Our cellular models natively express the BcrAbl oncogene and are either sensitive or have acquired resistance to well-characterized BcrAbl tyrosine kinase inhibitors. This approach measures IC50 values comparable to established methods of assessing drug potency, and its robustness indicates that it can be employed in drug discovery applications. This medium-throughput assay could bridge the gap between single target focused, high-throughput in vitro assays and lower-throughput cell-based follow-up experiments.
| Original language | English (US) |
|---|---|
| Article number | e0161748 |
| Journal | PloS one |
| Volume | 11 |
| Issue number | 9 |
| DOIs | |
| State | Published - Sep 2016 |
Bibliographical note
Funding Information:This work was supported by the National Institutes of Health through grants R00CA127161 and R21CA160129. The authors would like to thank Matthew Martin and Broxton Davis for assisting in experimental procedures.
Publisher Copyright:
Copyright © 2016 Ouellette et al.This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
