Roscovitine targets, protein kinases and pyridoxal kinase.
(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.
CNRS, Cell Cycle Group, UPS 2682 & UMR 2775, Station Biologique, BP 74, 29682 Roscoff cedex, Bretagne, France.
Stéphane
S
Bach
Marie
M
Knockaert
Jens
J
Reinhardt
Olivier
O
Lozach
Sophie
S
Schmitt
Blandine
B
Baratte
Marcel
M
Koken
Stephen P
SP
Coburn
Lin
L
Tang
Tao
T
Jiang
Dong-Cai
DC
Liang
Hervé
H
Galons
Jean-Francois
JF
Dierick
Lorenzo A
LA
Pinna
Flavio
F
Meggio
Frank
F
Totzke
Christoph
C
Schächtele
Andrea S
AS
Lerman
Amancio
A
Carnero
Yongqin
Y
Wan
Nathanael
N
Gray
Laurent
L
Meijer
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D015394:Molecular Structure; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011687:Purines / Q000737:chemistry* / Q000378:metabolism*; D011730:Pyridoxal / Q000378:metabolism; D011731:Pyridoxal Kinase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011732:Pyridoxal Phosphate / Q000378:metabolism; D051381:Rats; D000077546:Roscovitine; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014018:Tissue Distribution
D047428:Protein Kinase Inhibitors; D011687:Purines; D000077546:Roscovitine; D011730:Pyridoxal; D011732:Pyridoxal Phosphate; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D011731:Pyridoxal Kinase