Development of a novel fluorescent probe for fluorescence correlation spectroscopic detection of kinase inhibitors

Mitsuyasu Kawaguchi; Takuya Terai; Rei Utata; Miki Kato; Keiko Tsuganezawa; Akiko Tanaka; Hirotatsu Kojima; Takayoshi Okabe; Tetsuo Nagano

doi:10.1016/j.bmcl.2008.05.040

Development of a novel fluorescent probe for fluorescence correlation spectroscopic detection of kinase inhibitors

Bioorg Med Chem Lett. 2008 Jul 1;18(13):3752-5. doi: 10.1016/j.bmcl.2008.05.040. Epub 2008 May 16.

Authors

Mitsuyasu Kawaguchi¹, Takuya Terai, Rei Utata, Miki Kato, Keiko Tsuganezawa, Akiko Tanaka, Hirotatsu Kojima, Takayoshi Okabe, Tetsuo Nagano

Affiliation

¹ Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

PMID: 18524589
DOI: 10.1016/j.bmcl.2008.05.040

Abstract

We have developed a fluorescently labeled probe for high-throughput screening of kinase inhibitors using fluorescence correlation spectroscopy. With this probe, we have successfully evaluated the inhibitory activities of known inhibitors of a model kinase, ASK1. Because the probe contains a general kinase inhibitor, staurosporine, we believe that this homogeneous, high-throughput, and simple method can be applied to the inhibitor screening of other kinases as well.

MeSH terms

Chemistry, Pharmaceutical / methods
Crystallography, X-Ray / methods
Drug Evaluation, Preclinical
Fluorescence Polarization
Fluorescent Dyes / pharmacology*
Humans
Inhibitory Concentration 50
Kinetics
MAP Kinase Kinase Kinase 5 / metabolism
Models, Chemical
Molecular Conformation
Protein Binding
Protein Kinase Inhibitors / chemistry
Protein Kinase Inhibitors / pharmacology*
Spectrometry, Fluorescence / instrumentation*
Spectrometry, Fluorescence / methods*
Staurosporine / chemistry
Staurosporine / pharmacology

Substances

Fluorescent Dyes
Protein Kinase Inhibitors
MAP Kinase Kinase Kinase 5
Staurosporine