1H25

CDK2/Cyclin A in complex with an 11-residue recruitment peptide from retinoblastoma-associated protein

Summary for 1H25

• Entry DOI: 10.2210/pdb1h25/pdb
• Related: 1AD6 1AQ1 1B38 1B39 1BUH 1CKP 1DI8 1DM2 1E1V 1E1X 1E9H 1F5Q 1FIN 1FQ1 1FVT 1FVV 1G5S 1GH6 1GIH 1GII 1GIJ 1GUX 1GY3 1GZ8 1QMZ
• Descriptor: CELL DIVISION PROTEIN KINASE 2, CYCLIN A2, RETINOBLASTOMA-ASSOCIATED PROTEIN, ... (4 entities in total)
• Functional Keywords: transferase, cell cycle, protein kinase, cyclin, cdk2, recruitment, peptide specificity
• Biological source: HOMO SAPIENS (HUMAN); More
• Cellular location: Nucleus: P20248 P06400
• Total number of polymer chains: 5
• Total formula weight: 129785.34

Authors

Tews, I.,Cheng, K.Y.,Lowe, E.D.,Noble, M.E.M.,Brown, N.R.,Gul, S.,Gamblin, S.,Johnson, L.N. (deposition date: 2002-07-31, release date: 2003-02-01, Last modification date: 2024-10-16)

Primary citation

Lowe, E.D.,Tews, I.,Cheng, K.Y.,Brown, N.R.,Gul, S.,Noble, M.E.M.,Gamblin, S.,Johnson, L.N.
Specificity Determinants of Recruitment Peptides Bound to Phospho-Cdk2/Cyclin A
Biochemistry, 41:15625-, 2002

PubMed Abstract: Progression through S phase of the eukaryotic cell cycle is regulated by the action of the cyclin dependent protein kinase 2 (CDK2) in association with cyclin A. CDK2/cyclin A phosphorylates numerous substrates. Substrate specificity often employs a dual recognition strategy in which the sequence flanking the phospho-acceptor site (Ser.Pro.X.Arg/Lys) is recognized by CDK2, while the cyclin A component of the complex contains a hydrophobic site that binds Arg/Lys.X.Leu ("RXL" or "KXL") substrate recruitment motifs. To determine additional sequence specificity motifs around the RXL sequence, we have performed X-ray crystallographic studies at 2.3 A resolution and isothermal calorimetry measurements on complexes of phospho-CDK2/cyclin A with a recruitment peptide derived from E2F1 and with shorter 11-mer peptides from p53, pRb, p27, E2F1, and p107. The results show that the cyclin recruitment site accommodates a second hydrophobic residue either immediately C-terminal or next adjacent to the leucine of the "RXL" motif and that this site makes important contributions to the recruitment peptide recognition. The arginine of the RXL motif contacts a glutamate, Glu220, on the cyclin. In those substrates that contain a KXL motif, no ionic interactions are observed with the lysine. The sequences N-terminal to the "RXL" motif of the individual peptides show no conservation, but nevertheless make common contacts to the cyclin through main chain interactions. Thus, the recruitment site is able to recognize diverse but conformationally constrained target sequences. The observations have implications for the further identification of physiological substrates of CDK2/cyclin A and the design of specific inhibitors.

PubMed: 12501191
DOI: 10.1021/BI0268910

Experimental method

X-RAY DIFFRACTION (2.5 Å)