title: A TPX2 Proteomimetic Has Enhanced Affinity for Aurora-A Due to Hydrocarbon Stapling of a Helix.
abstract: Inhibition of protein kinases using ATP-competitive compounds is an important strategy in drug discovery. In contrast, the allosteric regulation of kinases through the disruption of protein-protein interactions has not been widely adopted, despite the potential for selective targeting. Aurora-A kinase regulates mitotic entry and mitotic spindle assembly and is a promising target for anticancer therapy. The microtubule-associated protein TPX2 activates Aurora-A through binding to two sites. Aurora-A recognition is mediated by two motifs within the first 43 residues of TPX2, connected by a flexible linker. To characterize the contributions of these three structural elements, we prepared a series of TPX2 proteomimetics and investigated their binding affinity for Aurora-A using isothermal titration calorimetry. A novel stapled TPX2 peptide was developed that has improved binding affinity for Aurora-A and mimics the function of TPX2 in activating Aurora-A's autophosphorylation. We conclude that the helical region of TPX2 folds upon binding Aurora-A, and that stabilization of this helix does not compromise Aurora-A activation. This study demonstrates that the preparation of these proteomimetics using modern synthesis methods is feasible and their biochemical evaluation demonstrates the power of proteomimetics as tool compounds for investigating PPIs involving intrinsically disordered regions of proteins.
authors: Department of Chemistry, University of Leicester , Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Yana K
authors: YK
authors: Rennie
authors: Department of Molecular and Cell Biology, University of Leicester , Lancaster Road, Leicester LE1 9HN, United Kingdom.
authors: Patrick J
authors: PJ
authors: McIntyre
authors: Department of Chemistry, University of Leicester , Lancaster Road, Leicester LE1 9HN, United Kingdom.|International Institute for Integrative Sleep Medicine, University of Tsukuba , Ibaraki 305-8575, Japan.
authors: Tito
authors: T
authors: Akindele
authors: Department of Molecular and Cell Biology, University of Leicester , Lancaster Road, Leicester LE1 9HN, United Kingdom.|Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds , Leeds, LS2 9JT, United Kingdom.
authors: Richard
authors: R
authors: Bayliss
authors: School of Chemistry, Joseph Black Building, University Avenue , Glasgow, G12 8QQ, United Kingdom.
authors: Andrew G
authors: AG
authors: Jamieson
mesh_terms: D000595:Amino Acid Sequence; D064096:Aurora Kinase A / Q000737:chemistry / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000378:metabolism*; D062105:Molecular Docking Simulation; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D060066:Protein Interaction Maps; D013816:Thermodynamics
chemical_list: D018797:Cell Cycle Proteins; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; C122167:TPX2 protein, human; D064096:Aurora Kinase A