Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

Click here to sign up for SAGE Journal Email Alerts today!

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Free Full Text (Free PDF) Free
Right arrow All Versions of this Article:
1087057107303323v1
12/6/809    most recent
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Web of Science (1)
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Ishii, T.
Right arrow Articles by Noumi, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ishii, T.
Right arrow Articles by Noumi, T.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

A Robust, Target-Driven, Cell-Based Assay for Checkpoint Kinase 1 Inhibitors

Tsuyoshi Ishii

GlaxoSmithKline K.K., Tsukuba-shi Ibaraki, Japan

Hiroshi Sootome

Bioscience Department GlaxoSmithKline K.K. 43 Wadai, Tsukuba-shi Ibaraki 300-4247, Japan, hiroshi_sootome{at}merck.com

Alastair J. King

GlaxoSmithKline, Upper Providence, Pennsylvania

Mikiya Suda

GlaxoSmithKline K.K., Tsukuba-shi Ibaraki, Japan, Molecular Medicine Research Laboratories, Drug Discovery Research, Astellas Pharma Inc., Tsukuba, Ibaraki, Japan

Nobuhiro Noro

GlaxoSmithKline K.K., Tsukuba-shi Ibaraki, Japan

Keizo Yamashita

GlaxoSmithKline K.K., Tsukuba-shi Ibaraki, Japan

Takato Noumi

GlaxoSmithKline K.K., Tsukuba-shi Ibaraki, Japan

Checkpoint kinase 1 (Chk1), a serine/threonine kinase, plays an important role in DNA damage checkpoint control and is an attractive target for cancer treatment. To develop a Chk1-specific cell-based assay, stable clones were established in which Chk1 kinase domain fused at its N-terminus with p53 through 4 tandem repeats of Gly-Gly-Gly-Gly-Ser was expressed in an inducible manner. Chk1 kinase specificity of the phosphorylation of fused p53 was confirmed by the experiments with a kinase-inactive Chk1. Only in the presence of an inducer molecule was phosphorylation of p53 at Ser-15 in the stable clones induced. Furthermore, its assay performance proved acceptable for high-throughput screening applications, judging from the Z' factor values (> 0.77). Finally, the cell-based assay thus established yielded structure-activity relationship data for a small set of test inhibitors of Chk1 within cells. Collectively, these results demonstrate that the established cell-based assay provides a novel and highly sensitive cellular platform for Chk1 inhibitor discovery. (Journal of Biomolecular Screening 2007:809-817)

Key Words: checkpoint kinase 1 • GeneSwitch system • target specific • cell-based assay

This version was published on September 1, 2007

Journal of Biomolecular Screening, Vol. 12, No. 6, 809-817 (2007)
DOI: 10.1177/1087057107303323
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?