Roscovitine Targets, Protein Kinases and Pyridoxal Kinase[boxs]

• Published in: The Journal of biological chemistry Vol. 280; no. 35; pp. 31208 - 31219
• Main Authors: Bach, Stéphane, Knockaert, Marie, Reinhardt, Jens, Lozach, Olivier, Schmitt, Sophie, Baratte, Blandine, Koken, Marcel, Coburn, Stephen P., Tang, Lin, Jiang, Tao, Liang, Dong-cai, Galons, Hervé, Dierick, Jean-Francois, Pinna, Lorenzo A., Meggio, Flavio, Totzke, Frank, Schächtele, Christoph, Lerman, Andrea S., Carnero, Amancio, Wan, Yongqin, Gray, Nathanael, Meijer, Laurent
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 02.09.2005; American Society for Biochemistry and Molecular Biology
• Subjects: Adenosine Triphosphate - metabolism; Amino Acid Sequence; Animals; Biochemistry, Molecular Biology; Cell Cycle - physiology; Cell Survival; Cells, Cultured; Chromatography, Affinity; Fibroblasts - cytology; Fibroblasts - physiology; Humans; Life Sciences; Mice; Mice, Knockout; Models, Molecular; Molecular Conformation; Molecular Sequence Data; Molecular Structure; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - metabolism; Protein Kinases - metabolism; Protein Structure, Tertiary; Purines - chemistry; Purines - metabolism; Pyridoxal - metabolism; Pyridoxal Kinase - antagonists & inhibitors; Pyridoxal Kinase - genetics; Pyridoxal Kinase - metabolism; Pyridoxal Phosphate - metabolism; Rats; Roscovitine; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tissue Distribution
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M500806200

(R)-Roscovitine (CYC202) is often referred to as a “selective inhibitor of cyclin-dependent kinases.” Besides its use as a biological tool in cell cycle, neuronal functions, and apoptosis studies, it is currently evaluated as a potential drug to treat cancers, neurodegenerative diseases, viral infections, and glomerulonephritis. We have investigated the selectivity of (R)-roscovitine using three different methods: 1) testing on a wide panel of purified kinases that, along with previously published data, now reaches 151 kinases; 2) identifying roscovitine-binding proteins from various tissue and cell types following their affinity chromatography purification on immobilized roscovitine; 3) investigating the effects of roscovitine on cells deprived of one of its targets, CDK2. Altogether, the results show that (R)-roscovitine is rather selective for CDKs, in fact most kinases are not affected. However, it binds an unexpected, non-protein kinase target, pyridoxal kinase, the enzyme responsible for phosphorylation and activation of vitamin B6. These results could help in interpreting the cellular actions of (R)-roscovitine but also in guiding the synthesis of more selective roscovitine analogs.