A novel quantitative kinase assay using bacterial surface display and flow cytometry

Henriques, Sónia Troeira, Thorstholm, Louise, Huang, Yen-Hua, Getz, Jennifer A., Daugherty, Patrick S. and Craik, David J. (2013) A novel quantitative kinase assay using bacterial surface display and flow cytometry. PloS One, 8 11: e80474.1-e80474.7. doi:10.1371/journal.pone.0080474

Author Henriques, Sónia Troeira
Thorstholm, Louise
Huang, Yen-Hua
Getz, Jennifer A.
Daugherty, Patrick S.
Craik, David J.
Title A novel quantitative kinase assay using bacterial surface display and flow cytometry
Journal name PloS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2013-11-15
Sub-type Article (original research)
DOI 10.1371/journal.pone.0080474
Open Access Status DOI
Volume 8
Issue 11
Start page e80474.1
End page e80474.7
Total pages 7
Editor Emilio Hirsch
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Collection year 2014
Language eng
Formatted abstract
The inhibition of tyrosine kinases is a successful approach for the treatment of cancers and the discovery of kinase inhibitor drugs is the focus of numerous academic and pharmaceutical laboratories. With this goal in mind, several strategies have been developed to measure kinase activity and to screen novel tyrosine kinase inhibitors. Nevertheless, a general non-radioactive and inexpensive approach, easy to implement and adapt to a range of applications, is still missing. Herein, using Bcr-Abl tyrosine kinase, an oncogenic target and a model protein for cancer studies, we describe a novel cost-effective high-throughput screening kinase assay. In this approach, named the BacKin assay, substrates displayed on a Bacterial cell surface are incubated with Kinase and their phosphorylation is examined and quantified by flow cytometry. This approach has several advantages over existing approaches, as using bacteria (i.e. Escherichia coli) to display peptide substrates provides a self renewing solid support that does not require laborious chemical strategies. Here we show that the BacKin approach can be used for kinetic and mechanistic studies, as well as a platform to characterize and identify small-molecule or peptide-based kinase inhibitors with potential applications in drug development.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
Institute for Molecular Bioscience - Publications
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Citation counts: TR Web of Science Citation Count  Cited 5 times in Thomson Reuters Web of Science Article | Citations
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Created: Tue, 10 Dec 2013, 11:54:56 EST by Susan Allen on behalf of Institute for Molecular Bioscience