5EW9
Crystal Structure of Aurora A Kinase Domain Bound to MK-5108
Summary for 5EW9
| Entry DOI | 10.2210/pdb5ew9/pdb |
| Descriptor | Aurora kinase A, 4-(3-chloranyl-2-fluoranyl-phenoxy)-1-[[6-(1,3-thiazol-2-ylamino)pyridin-2-yl]methyl]cyclohexane-1-carboxylic acid (3 entities in total) |
| Functional Keywords | transferase, protein kinase, inhibitor, transferase-transferase inhibitor complex, transferase/transferase inhibitor |
| Biological source | Homo sapiens (Human) |
| Cellular location | Cytoplasm, cytoskeleton, microtubule organizing center, centrosome : O14965 |
| Total number of polymer chains | 1 |
| Total formula weight | 31998.99 |
| Authors | Shiau, A.K.,Motamedi, A. (deposition date: 2015-11-20, release date: 2016-01-20, Last modification date: 2024-10-23) |
| Primary citation | de Groot, C.O.,Hsia, J.E.,Anzola, J.V.,Motamedi, A.,Yoon, M.,Wong, Y.L.,Jenkins, D.,Lee, H.J.,Martinez, M.B.,Davis, R.L.,Gahman, T.C.,Desai, A.,Shiau, A.K. A Cell Biologist's Field Guide to Aurora Kinase Inhibitors. Front Oncol, 5:285-285, 2015 Cited by PubMed Abstract: Aurora kinases are essential for cell division and are frequently misregulated in human cancers. Based on their potential as cancer therapeutics, a plethora of small molecule Aurora kinase inhibitors have been developed, with a subset having been adopted as tools in cell biology. Here, we fill a gap in the characterization of Aurora kinase inhibitors by using biochemical and cell-based assays to systematically profile a panel of 10 commercially available compounds with reported selectivity for Aurora A (MLN8054, MLN8237, MK-5108, MK-8745, Genentech Aurora Inhibitor 1), Aurora B (Hesperadin, ZM447439, AZD1152-HQPA, GSK1070916), or Aurora A/B (VX-680). We quantify the in vitro effect of each inhibitor on the activity of Aurora A alone, as well as Aurora A and Aurora B bound to fragments of their activators, TPX2 and INCENP, respectively. We also report kinome profiling results for a subset of these compounds to highlight potential off-target effects. In a cellular context, we demonstrate that immunofluorescence-based detection of LATS2 and histone H3 phospho-epitopes provides a facile and reliable means to assess potency and specificity of Aurora A versus Aurora B inhibition, and that G2 duration measured in a live imaging assay is a specific readout of Aurora A activity. Our analysis also highlights variation between HeLa, U2OS, and hTERT-RPE1 cells that impacts selective Aurora A inhibition. For Aurora B, all four tested compounds exhibit excellent selectivity and do not significantly inhibit Aurora A at effective doses. For Aurora A, MK-5108 and MK-8745 are significantly more selective than the commonly used inhibitors MLN8054 and MLN8237. A crystal structure of an Aurora A/MK-5108 complex that we determined suggests the chemical basis for this higher specificity. Taken together, our quantitative biochemical and cell-based analyses indicate that AZD1152-HQPA and MK-8745 are the best current tools for selectively inhibiting Aurora B and Aurora A, respectively. However, MK-8745 is not nearly as ideal as AZD1152-HQPA in that it requires high concentrations to achieve full inhibition in a cellular context, indicating a need for more potent Aurora A-selective inhibitors. We conclude with a set of "good practice" guidelines for the use of Aurora inhibitors in cell biology experiments. PubMed: 26732741DOI: 10.3389/fonc.2015.00285 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.181 Å) |
Structure validation
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