Homogeneous and nonradioactive high-throughput screening platform for the characterization of kinase inhibitors in cell lysates

Détails

ID Serval
serval:BIB_95855612ABCB
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Homogeneous and nonradioactive high-throughput screening platform for the characterization of kinase inhibitors in cell lysates
Périodique
Journal of Biomolecular Screening
Auteur(s)
Guenat S., Rouleau N., Bielmann C., Bedard J., Maurer F., Allaman-Pillet N., Nicod P., Bielefeld-Sevigny M., Beckmann J. S., Bonny C., Bosse R., Roduit R.
ISSN
1087-0571 (Print)
Statut éditorial
Publié
Date de publication
12/2006
Peer-reviewed
Oui
Volume
11
Numéro
8
Pages
1015-26
Langue
anglais
Notes
Evaluation Studies
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Dec
Résumé
Protein kinases are directly implicated in many human diseases; therefore, kinase inhibitors show great promises as new therapeutic drugs. In an effort to facilitate the screening and the characterization of kinase inhibitors, a novel application of the AlphaScreen technology was developed to monitor JNK activity from (1) purified kinase preparations and (2) endogenous kinase from cell lysates preactivated with different cytokines. The authors confirmed that both adenosine triphosphate (ATP) competitive as well as peptide-based JNK inhibitors were able to block the activity of both recombinant and HepG2 endogenous JNK activity. Using the same luminescence technique adapted for binding studies, the authors characterized peptide inhibitor mechanisms by measuring the binding affinity of the inhibitors for JNK. Because of the versatility of the technology, this cell-based JNK kinase assay could be adapted to other kinases and would represent a powerful tool to evaluate endogenous kinase activity and test a large number of potential inhibitors in a more physiologically relevant environment.
Mots-clé
Binding Sites Binding, Competitive Cell Line Combinatorial Chemistry Techniques/*methods Dose-Response Relationship, Drug Drug Evaluation, Preclinical/*methods Enzyme Inhibitors/*chemistry Humans MAP Kinase Kinase 4/metabolism Protein Kinases/*metabolism
Pubmed
Web of science
Création de la notice
25/01/2008 17:18
Dernière modification de la notice
03/03/2018 19:40
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