--- _id: '3983' abstract: - lang: eng text: Cdc25 phosphatases are key activators of the eukaryotic cell cycle and compelling anticancer targets because their overexpression has been associated with numerous cancers. However, drug discovery targeting these phosphatases has been hampered by the lack of structural information about how Cdc25s interact with their native protein substrates, the cyclin-dependent kinases. Herein, we predict a docked orientation for Cdc25B with its Cdk2-pTpY-CycA protein substrate by a rigid-body docking method and refine the docked models with full-scale molecular dynamics simulations and minimization. We validate the stable ensemble structure experimentally by a variety of in vitro and in vivo techniques. Specifically, we compare our model with a crystal structure of the substrate-trapping mutant of Cdc25B. We identify and validate in vivo a novel hot-spot residue on Cdc25B (Arg492) that plays a central role in protein substrate recognition. We identify a hot-spot residue on the Substrate Cdk2 (Asp206) and confirm its interaction with hot-spot residues on Cdc25 using hot-spot swapping and double mutant cycles to derive interaction energies. Our experimentally validated model is consistent with previous studies of Cdk2 and its interaction partners and initiates the opportunity for drug discovery of inhibitors that target the remote binding sites of this protein-protein interaction. author: - first_name: Jungsan full_name: Sohn, Jungsan last_name: Sohn - first_name: Jerry full_name: Parks, Jerry M last_name: Parks - first_name: Gregory full_name: Buhrman, Gregory last_name: Buhrman - first_name: Paul full_name: Brown, Paul last_name: Brown - first_name: Kolbrun full_name: Kristjánsdóttir, Kolbrun last_name: Kristjánsdóttir - first_name: Alexias full_name: Safi, Alexias last_name: Safi - first_name: Herbert full_name: Herbert Edelsbrunner id: 3FB178DA-F248-11E8-B48F-1D18A9856A87 last_name: Edelsbrunner orcid: 0000-0002-9823-6833 - first_name: Weitao full_name: Yang, Weitao T last_name: Yang - first_name: Johannes full_name: Rudolph, Johannes last_name: Rudolph citation: ama: Sohn J, Parks J, Buhrman G, et al. Experimental validation of the docking orientation of Cdc25 with its Cdk2-CycA protein substrate. Biochemistry. 2005;44(50):16563-16573. doi:10.1021/bi0516879 apa: Sohn, J., Parks, J., Buhrman, G., Brown, P., Kristjánsdóttir, K., Safi, A., … Rudolph, J. (2005). Experimental validation of the docking orientation of Cdc25 with its Cdk2-CycA protein substrate. Biochemistry. ACS. https://doi.org/10.1021/bi0516879 chicago: Sohn, Jungsan, Jerry Parks, Gregory Buhrman, Paul Brown, Kolbrun Kristjánsdóttir, Alexias Safi, Herbert Edelsbrunner, Weitao Yang, and Johannes Rudolph. “Experimental Validation of the Docking Orientation of Cdc25 with Its Cdk2-CycA Protein Substrate.” Biochemistry. ACS, 2005. https://doi.org/10.1021/bi0516879. ieee: J. Sohn et al., “Experimental validation of the docking orientation of Cdc25 with its Cdk2-CycA protein substrate,” Biochemistry, vol. 44, no. 50. ACS, pp. 16563–16573, 2005. ista: Sohn J, Parks J, Buhrman G, Brown P, Kristjánsdóttir K, Safi A, Edelsbrunner H, Yang W, Rudolph J. 2005. Experimental validation of the docking orientation of Cdc25 with its Cdk2-CycA protein substrate. Biochemistry. 44(50), 16563–16573. mla: Sohn, Jungsan, et al. “Experimental Validation of the Docking Orientation of Cdc25 with Its Cdk2-CycA Protein Substrate.” Biochemistry, vol. 44, no. 50, ACS, 2005, pp. 16563–73, doi:10.1021/bi0516879. short: J. Sohn, J. Parks, G. Buhrman, P. Brown, K. Kristjánsdóttir, A. Safi, H. Edelsbrunner, W. Yang, J. Rudolph, Biochemistry 44 (2005) 16563–16573. date_created: 2018-12-11T12:06:16Z date_published: 2005-11-24T00:00:00Z date_updated: 2021-01-12T07:53:39Z day: '24' doi: 10.1021/bi0516879 extern: 1 intvolume: ' 44' issue: '50' month: '11' page: 16563 - 16573 publication: Biochemistry publication_status: published publisher: ACS publist_id: '2144' quality_controlled: 0 status: public title: Experimental validation of the docking orientation of Cdc25 with its Cdk2-CycA protein substrate type: journal_article volume: 44 year: '2005' ...