The Role of the Phospho-CDK2/Cyclin A Recruitment Site in Substrate Recognition
作者： Kin-Yip Cheng , Martin E. M. Noble , Vicky Skamnaki , Nick R. Brown , Ed D. Lowe
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摘要: Phospho-CDK2/cyclin A, a kinase that is active in cell cycle S phase, contains an RXL substrate recognition site over 40 A from the catalytic site. The role of this recruitment site, which enhances affinity and efficiency, has been investigated using peptides derived natural substrates, namely CDC6 p107, bispeptide inhibitor gamma-phosphate ATP covalently attached by linker to peptide. X-ray studies with 30-residue peptide complex pCDK2/cyclin showed binding dodecamer at heptapeptide but no density for linking 11 residues. Kinetic established had 18-fold lower Km compared effect required be linked modified two alternative conformations was potent competitive both activity against (Ki = 0.83 nm) less effective RXL-containing substrates. We discuss how localization (KD 0.4 microm) leads increased efficiency design inhibitor. notion flexible between sites, must have more than minimal number residues, provides explanation discrimination different
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