e-Science logo Nesc logo
About NeSC
e-Science Institute
e-Science Hub
e-Science Events
Presentations & Lectures
Technical Papers
Global Grid Links
UK e-Science Centres
UK e-Science Teams
Career Opportunities
Bibliographic Database


Paper ID: 2736

Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors
M Knockaerta,N Grayb

Appeared in: Chemistry and Biology
Page Numbers:411-422
Publisher: N/A
Year: 2000
Contributing Organisation(s): Station Biologique de Roscof, Novartis Institute for Functional Genomics, The Scripps Research Institute, Museum d’Histoire Naturelle, Wellcome Centre for Molecular Parasitology,Unité INSERM U511
Field of Science: Medicine

URL: http://carlin.lib.ed.ac.uk:2068/science?_ob=ArticleURL&_udi=B6VRP-40H57BK-8&_user=809099&_coverDate=06%2F01%2F2000&_alid=624645184&_rdoc=1&_fmt=full&_

Abstract: Background: Chemical inhibitors of cyclin-dependent kinases (CDKs) have great therapeutic potential against various proliferative and neurodegenerative disorders. Olomoucine, a 2,6,9-trisubstituted purine, has been optimized for activity against CDK1/cyclin B by combinatorial and medicinal chemistry efforts to yield the purvalanol inhibitors. Although many studies support the action of purvalanols against CDKs, the actual intracellular targets of 2,6,9-trisubstituted purines remain unverified. Results: To address this issue, purvalanol B (95) and an N6-methylated, CDK-inactive derivative (95M) were immobilized on an agarose matrix. Extracts from a diverse collection of cell types and organisms were screened for proteins binding purvalanol B. In addition to validating CDKs as intracellular targets, a variety of unexpected protein kinases were recovered from the 95 matrix. Casein kinase 1 (CK1) was identified as a principal 95 matrix binding protein in Plasmodium falciparum, Leishmania mexicana, Toxoplasma gondii and Trypanosoma cruzi. Purvalanol compounds also inhibit the proliferation of these parasites, suggesting that CK1 is a valuable target for further screening with 2,6,9-trisubstituted purine libraries. Conclusions: That a simple batchwise affinity chromatography approach using two purine derivatives facilitated isolation of a small set of highly purified kinases suggests that this could be a general method for identifying intracellular targets relevant to a particular class of ligands. This method allows a close correlation to be established between the pattern of proteins bound to a small family of related compounds and the pattern of cellular responses to these compounds.

Keywords: asein kinase 1; Cyclin-dependent kinases; Erk; Malaria; Purine



Last Updated: 22 Jun 12 11:02
This is an archived website, preserved and hosted by the School of Physics and Astronomy at the University of Edinburgh. The School of Physics and Astronomy takes no responsibility for the content, accuracy or freshness of this website. Please email webmaster [at] ph [dot] ed [dot] ac [dot] uk for enquiries about this archive.