Design of a Novel Class of Peptide Inhibitors of Cyclin-dependent Kinase/Cyclin Activation

• Published in: The Journal of biological chemistry Vol. 280; no. 14; pp. 13793 - 13800
• Main Authors: Gondeau, Claire, Gerbal-Chaloin, Sabine, Bello, Paul, Aldrian-Herrada, Gudrun, Morris, May C., Divita, Gilles
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 08.04.2005; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Antineoplastic Agents - chemistry; Antineoplastic Agents - metabolism; Biochemistry, Molecular Biology; CDC2-CDC28 Kinases - antagonists & inhibitors; CDC2-CDC28 Kinases - chemistry; CDC2-CDC28 Kinases - metabolism; Cell Cycle - physiology; Cell Line, Tumor; Cyclin A - chemistry; Cyclin A - genetics; Cyclin A - metabolism; Cyclin-Dependent Kinase 2; Drug Design; Enzyme Activation; Enzyme Inhibitors - metabolism; Enzymz Inhibitors / metabolism; Gene Products, tat - genetics; Gene Products, tat - metabolism; Humans; Life Sciences; Macromolecular Substances; Models, Molecular; Molecular Sequence Data; Peptides - chemical synthesis; Peptides - chemistry; Peptides - genetics; Peptides - metabolism; Protein Binding; Protein Conformation; Protein Subunits - chemistry; Protein Subunits - genetics; Protein Subunits - metabolism; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Sequence Alignment; Rb; CD; N-(9-fluorenyl)methoxycarbonyl; retinoblastoma; glutathione S-transferase; GST; Fmoc; CDK; circular dichroism; cyclin-dependent kinase; cyclin binding motif; CBM
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M413690200

Cyclin dependent kinases (CDKs) are key regulators of the cell cycle progression and therefore constitute excellent targets for the design of anticancer agents. Most of the inhibitors identified to date inhibit kinase activity by interfering with the ATP-binding site of CDKs. We recently proposed that the protein/protein interface and conformational changes required in the molecular mechanism of CDK2-cyclin A activation were potential targets for the design of specific inhibitors of cell cycle progression. To this aim, we have designed and characterized a small peptide, termed C4, derived from amino acids 285–306 in the α5 helix of cyclin A. We demonstrate that this peptide does not interfere with complex formation but forms stable complexes with CDK2-cyclin A. The C4 peptide significantly inhibits kinase activity of complexes harboring CDK2 in a competitive fashion with respect to substrates but does not behave as an ATP antagonist. Moreover, when coupled with the protein transduction domain of Tat, the C4 peptide blocks the proliferation of tumor cell lines, thereby constituting a potent lead for the development of specific CDK-cyclin inhibitors.